U.S. patent application number 12/131923 was filed with the patent office on 2009-09-17 for abuse-resistant amphetamine prodrugs.
This patent application is currently assigned to SHIRE LLC. Invention is credited to Barney Bishop, Suma Krishnan, Christopher Lauderback, Travis Mickle, James Scott Moncrief, Robert Oberlender, Bernhard J. Paul, Thomas Piccariello, Christopher A. Verbicky.
Application Number | 20090234002 12/131923 |
Document ID | / |
Family ID | 46331900 |
Filed Date | 2009-09-17 |
United States Patent
Application |
20090234002 |
Kind Code |
A1 |
Mickle; Travis ; et
al. |
September 17, 2009 |
ABUSE-RESISTANT AMPHETAMINE PRODRUGS
Abstract
The invention describes compounds, compositions, and methods of
using the same comprising a chemical moiety covalently attached to
amphetamine. These compounds and compositions are useful for
reducing or preventing abuse and overdose of amphetamine. These
compounds and compositions find particular use in providing an
abuse-resistant alternative treatment for certain disorders, such
as attention deficit hyperactivity disorder (ADHD), ADD,
narcolepsy, and obesity. Oral bioavailability of amphetamine is
maintained at therapeutically useful doses. At higher doses
bioavailability is substantially reduced, thereby providing a
method of reducing oral abuse liability. Further, compounds and
compositions of the invention decrease the bioavailability of
amphetamine by parenteral routes, such as intravenous or intranasal
administration, further limiting their abuse liability.
Inventors: |
Mickle; Travis; (Coralville,
IA) ; Krishnan; Suma; (Belvedere, CA) ;
Bishop; Barney; (Annandale, VA) ; Lauderback;
Christopher; (Blacksburg, VA) ; Moncrief; James
Scott; (Christiansburg, VA) ; Oberlender; Robert;
(Blacksburg, VA) ; Piccariello; Thomas;
(Blacksburg, VA) ; Paul; Bernhard J.; (Lexington,
MA) ; Verbicky; Christopher A.; (Broadalbin,
NY) |
Correspondence
Address: |
DARBY & DARBY P.C.
P.O. BOX 770, Church Street Station
New York
NY
10008-0770
US
|
Assignee: |
SHIRE LLC
Florence
KY
|
Family ID: |
46331900 |
Appl. No.: |
12/131923 |
Filed: |
June 2, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11400304 |
Apr 10, 2006 |
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12131923 |
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|
10857619 |
Jun 1, 2004 |
7223735 |
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11400304 |
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10858526 |
Jun 1, 2004 |
7105486 |
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10857619 |
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PCT/US03/05525 |
Feb 24, 2003 |
|
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10858526 |
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60358368 |
Feb 22, 2002 |
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60362082 |
Mar 7, 2002 |
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60473929 |
May 29, 2003 |
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60567801 |
May 5, 2004 |
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60669385 |
Apr 8, 2005 |
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60669386 |
Apr 8, 2005 |
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60681170 |
May 16, 2005 |
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60756548 |
Jan 6, 2006 |
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60759958 |
Jan 19, 2006 |
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Current U.S.
Class: |
514/483 ;
560/16 |
Current CPC
Class: |
A61K 31/27 20130101;
A61K 47/542 20170801; C07C 237/06 20130101; A61K 31/165 20130101;
Y10T 436/173845 20150115; Y10S 436/901 20130101 |
Class at
Publication: |
514/483 ;
560/16 |
International
Class: |
A61K 31/27 20060101
A61K031/27; C07C 271/08 20060101 C07C271/08 |
Claims
1. Crystalline lisdexamphetamine dimesylate.
2. The crystalline lisdexamphetamine dimesylate of claim 1, wherein
the crystalline lisdexamphetamine dimesylate exhibits an X-ray
powder diffraction pattern substantially as shown in FIG. 77.
3. The crystalline lisdexamphetamine dimesylate of claim 1, wherein
the crystalline lisdexamphetamine dimesylate exhibits an X-ray
powder diffraction pattern having at least one peak in degrees 2
.+-.0.2 2 selected from 4.5, 9.0, 12.0, 15.7, and 16.3.
4. The crystalline lisdexamphetamine dimesylate of claim 3, wherein
the dimesylate exhibits an X-ray powder diffraction pattern having
at least two peaks in degrees 2.THETA..+-.0.2 2.THETA. selected
from 4.5, 9.0, 12.0, 15.7, and 16.3.
5. The crystalline lisdexamphetamine dimesylate of claim 4, wherein
the dimesylate exhibits an X-ray powder diffraction pattern having
peaks in degrees 2.THETA..+-.0.2 2.THETA. at 4.5, 9.0, 12.0 and
15.7.
6. Crystalline lisdexamphetamine dimesylate exhibiting an X-ray
powder diffraction pattern having at least one peak in degrees
2.THETA..+-.0.2 2.THETA. selected from 4.5, 9.0, 12.0, 15.7, and
16.3.
7. The crystalline lisdexamphetamine dimesylate of claim 1, wherein
the crystalline lisdexamphetamine dimesylate has a melting point
onset as determined by differential scanning calorimetry at about
194.7.degree. C.
8. The crystalline lisdexamphetamine dimesylate of claim 1, wherein
the crystalline lisdexamphetamine dimesylate exhibits a single
crystal X-ray crystallographic analysis at 150 K with crystal
parameters that are approximately equal to the following:
TABLE-US-00088 Parameter Value Space group P2.sub.1/a Cell
Dimensions a (.ANG.) 10.25 b (.ANG.) 11.28 c (.ANG.) 19.35 ( )
94.12 Volume (.ANG..sup.3) 2232.1 Z (Molecules/unit cell) 4 Density
(g/cm.sup.3) 1.359
9. Crystalline lisdexamphetamine dimesylate particles having at
least one of: (a) a d.sub.10 ranging from about 1 to about 10
.mu.m; (b) a d.sub.50 ranging from about 8 to about 40 .mu.m; and
(c) a d.sub.90 particle size ranging from about 25 to about 90
.mu.m.
10. The crystalline lisdexamphetamine dimesylate particles of claim
9, wherein the crystalline lisdexamphetamine dimesylate has a
d.sub.90 particle size ranging from about 25 to about 90 .mu.m.
11. The crystalline lisdexamphetamine dimesylate particles of claim
9, wherein the crystalline lisdexamphetamine dimesylate has a
d.sub.50 ranging from about 8 to about 40 .mu.m.
12. (canceled)
13. A pharmaceutical composition comprising crystalline
lisdexamphetamine dimesylate of any one of claim 1 and a
pharmaceutically acceptable additive.
14. The pharmaceutical composition of claim 13, wherein the
pharmaceutical composition comprises at least about 95% by weight
of crystalline lisdexamphetamine dimesylate, based upon 100% total
weight of lisdexamphetamine dimesylate in the pharmaceutical
composition.
15. A method of treating attention deficit hyperactivity disorder
in a patient in need thereof, comprising the step of orally
administering the crystalline lisdexamphetamine dimesylate of claim
1.
16.-67. (canceled)
Description
CROSS REFERENCE RELATED APPLICATIONS
[0001] This application is a continuation-in-part of U.S.
application Ser. No. 11/400,304, filed Apr. 10, 2006, which in
turn, (a) claims benefit under 35 U.S.C. 119(e) to U.S. Provisional
Application Nos. 60/669,385 filed Apr. 8, 2005, 60/669,386 filed
Apr. 8, 2005, 60/681,170 filed May 16, 2005, 60/756,548 filed Jan.
6, 2006, and 60/759,958 filed Jan. 19, 2006; (b) is a
continuation-in-part of U.S. application Ser. No. 10/857,619 filed
Jun. 1, 2004, which claims the benefit of under 35 U.S.C. 119(e) to
U.S. Provisional Application Nos. 60/473,929 filed May 29, 2003 and
60/567,801 filed May 5, 2004; and (c) is a continuation-in-part of
U.S. application Ser. No. 10/858,526 filed Jun. 1, 2004, which, in
turn, is a continuation-in-part of international application
PCT/US03/05525 filed Feb. 24, 2003, which claims priority to U.S.
Provisional Application Nos. 60/358,368 filed Feb. 22, 2002 and
60/362,082 filed Mar. 7, 2002; application Ser. No. 10/858,526 also
claims benefit under 35 U.S.C. 119(e) to U.S. Provisional
Application Nos. 60/473,929 filed May 29, 2003 and 60/567,801 filed
May 5, 2004. All of the above-identified applications are hereby
incorporated by reference in their entirety.
FIELD OF THE INVENTION
[0002] The invention relates to amphetamine compounds, more
particularly to amphetamine prodrugs comprising amphetamine
covalently bound to a chemical moiety. The invention also relates
to pharmaceutical compositions comprising the amphetamine
compounds, and to methods of manufacturing, delivering, and using
the amphetamine compounds. The invention further relates to a
crystalline form of the amphetamine prodrug
(l)-lysine-(d)-amphetamine dimesylate.
BACKGROUND OF THE INVENTION
[0003] Amphetamines stimulate the central nervous system (CNS) and
have been used medicinally to treat various disorders including
attention deficit hyperactivity disorder (ADHD), obesity, and
narcolepsy. In children with ADHD, potent CNS stimulants have been
used for several decades as a drug treatment given either alone or
as an adjunct to behavioral therapy. While methylphenidate
(Ritalin.RTM.) has been the most frequently prescribed stimulant,
the prototype of the class, amphetamine (alpha-methyl
phenethylamine) has been used all along and increasingly so in
recent years. (Bradley C, Bowen M, "Amphetamine (benzedrine)
therapy of children's behavior disorders." American Journal of
Orthopsychiatry 11: 92-103 (1941).
[0004] Because of their stimulating effects, amphetamines,
including amphetamine derivatives and analogs, are subject to
abuse. A user can become dependent over time on these drugs and
their physical and psychological effects, even when the drugs are
used for legitimate therapeutic purposes. Legitimate amphetamine
users that develop drug tolerances are especially susceptible to
becoming accidental addicts as they increase dosing in order to
counteract their increased tolerance of the prescribed drugs.
Additionally, it is possible for individuals to inappropriately
self-administer higher than prescribed quantities of the drug or to
alter either the product or the route of administration (e.g.,
inhalation (snorting), injection, and smoking), potentially
resulting in immediate release of the active drug in quantities
larger than prescribed. When taken at higher than prescribed doses,
amphetamines can cause temporary feelings of exhilaration and
increased energy and mental alertness.
[0005] Recent developments in the abuse of prescription drug
products increasingly raise concerns about the abuse of amphetamine
prescribed for ADHD. The National Survey on Drug Use and Health
(NSDUH), estimates that in 2003, 1.2 million Americans aged 12 and
older abused stimulants, such as amphetamines. The high abuse
potential has earned amphetamines Schedule II status according to
the Controlled Substances Act (CSA). Schedule II classification is
reserved for those drugs that have accepted medical use but have
the highest potential for abuse.
[0006] Extended release formulations of amphetamines, e.g.,
Adderall XR.RTM., have an increased abuse liability relative to the
single dose tablets because each tablet of the sustained release
formulation contains a higher concentration of amphetamine. It may
be possible for substance abusers to obtain a high dose of
amphetamine with rapid onset by crushing the tablets into powder
and snorting it or by dissolving the powder in water and injecting
it. Sustained release formulations may also provide uneven
release.
[0007] Additional information about amphetamines and amphetamine
abuse can be found in U.S. Publication No. 2005/0054561 (U.S. Ser.
No. 10/858,526).
[0008] The need exists for additional amphetamine compounds,
especially abuse resistant amphetamine compounds. Further, the need
exists for amphetamine pharmaceutical compositions that provide
sustained release and sustained therapeutic effect.
SUMMARY OF THE INVENTION
[0009] The present invention provides amphetamine prodrugs, and
salts thereof, comprising amphetamine covalently bound to a
chemical moiety. A preferred amphetamine prodrug is
(l)-lysine-(d)-amphetamine dimesylate (also known as
lisdexamphetamine dimesylate). Lisdexamphetamine dimesylate can be
in crystalline form, and can be anhydrous (i.e., crystalline
anhydrous lisdexamphetamine dimesylate).
[0010] One embodiment of the invention is crystalline
lisdexamphetamine dimesylate which exhibits an X-ray powder
diffraction (XRPD) pattern having at least one peak in degrees
2.THETA..+-.0.2.degree. 2.THETA. selected from 4.5, 9.0, 12.0,
15.7, and 16.3. In another embodiment, the crystalline
lisdexamphetamine dimesylate exhibits 2, 3, 4, or all of the
aforementioned peaks. The crystalline lisdexamphetamine dimesylate
can exhibit an XRPD substantially as shown in FIG. 77.
[0011] Another embodiment of the invention is crystalline
lisdexamphetamine dimesylate having a melting point onset as
determined by differential scanning calorimetry (DSC) at about
194.7.degree. C.
[0012] Yet another embodiment is crystalline lisdexamphetamine
dimesylate that exhibits a single crystal X-ray crystallographic
analysis at 150 K with crystal parameters that are approximately
equal to the following:
TABLE-US-00001 Parameter Value Space group P2.sub.1/a Cell
Dimensions a (.ANG.) 10.25 b (.ANG.) 11.28 c (.ANG.) 19.35 .beta.
(.degree.) 94.12 Volume (.ANG..sup.3) 2232.1 Z (Molecules/unit
cell) 4 Density (g/cm.sup.3) 1.359
[0013] Still, another embodiment is crystalline lisdexamphetamine
dimesylate having at least one of a d.sub.10 ranging from about 1
to about 10 .mu.m; a d.sub.50 ranging from about 8 to about 40
.mu.m; and a d.sub.90 particle size ranging from about 25 to about
90 .mu.m.
[0014] The crystalline lisdexamphetamine dimesylate can be orally
administered to treat attention deficit hyperactivity disorder as
well as the other disorders described herein. For example, one
embodiment is a method of treating attention deficit hyperactivity
disorder in a patient (e.g., a child or adult) in need thereof by
administering an effective amount of the crystalline
lisdexamphetamine dimesylate.
[0015] In yet another aspect of the invention, a method for
preparing crystalline lisdexamphetamine dimesylate is provided. The
method entails reacting a di-amine protected l-lysine-d-amphetamine
with methane sulfonic acid to produce lisdexamphetamine
dimesylate.
[0016] Another embodiment of the invention is directed to a method
of reducing patient to patient variability of amphetamine levels
among a group of patients. The method entails daily (preferably
once daily) oral administration to each patient in the group of a
prodrug of amphetamine or a pharmaceutically acceptable salt of the
prodrug, wherein the amphetamine is covalently bound to a peptide
comprising 1 to 10 amino acids. In one preferred embodiment, a
pharmaceutical composition consisting essentially of the prodrug of
amphetamine or a pharmaceutically acceptable salt thereof is
administered. A preferred prodrug is L-lysine-d-amphetamine and
pharmaceutically acceptable salts thereof, such as
L-lysine-d-amphetamine dimesylate.
[0017] Yet another embodiment of the invention is a method of
treating obesity, cancer related fatigue, excessive daytime
sleepiness in patients suffering from obstructive sleep apnea, or
narcolepsy, in a patient in need thereof by administering an
effective amount of the aforementioned amphetamine prodrug or a
pharmaceutically acceptable salt thereof or a pharmaceutical
composition containing it. According to one preferred embodiment,
the prodrug or pharmaceutically acceptable salt thereof is orally
administered once daily. A preferred prodrug is
L-lysine-d-amphetamine, and a preferred salt of the prodrug is
L-lysine-d-amphetamine dimesylate.
[0018] Yet another embodiment of the invention is a method of
treating depression or a depressive disorder (such as major
depressive disorder) in a patient in need thereof by administering
an effective amount of (a) the aforementioned amphetamine prodrug
or a pharmaceutically acceptable salt thereof or a pharmaceutical
composition containing it and (b) an antidepressant other than
mirtazapine.
[0019] Yet another embodiment is a method of treating ADHD in a
patient in need thereof while minimizing the risk of adverse side
effects due to the mode of treatment, comprising:
(a) marketing to doctors an oral pharmaceutical composition
comprising L-lysine-d-amphetamine or a pharmaceutically acceptable
salt thereof for the treatment of ADHD and as having a lower
interpatient variation than an oral formulation containing
d-amphetamine; (b) diagnosing a patient as suffering from ADHD by a
doctor; (c) prescribing the oral pharmaceutical composition
comprising L-lysine-d-amphetamine or a pharmaceutically acceptable
salt thereof to the patient by the doctor in response to the
marketing of the oral pharmaceutical composition and the diagnosis
of the patient; and (d) orally administering the prescribed oral
pharmaceutical composition to the patient. The term "marketing"
refers to the act or process of selling a product, including, but
not limited to, any offer for sale or sale of a product, including
advertising The term "advertising" refers to notifying, informing,
and/or apprising one or more individuals of information (e.g., the
efficacy of a pharmaceutical product for treating or reducing an
indication), such as by mass media, including, but not limited to,
newspaper, magazine, and internet advertisements, television
commercials, and billboard signs. The term "advertising" as used
herein also includes including a statement that the pharmaceutical
product can treat the indication(s) in the labeling (e.g., ADHD)
for the pharmaceutical product.
[0020] Yet another embodiment is a method for treating a patient
suffering from ADHD comprising the steps of:
(a) providing to the marketplace an oral pharmaceutical composition
comprising L-lysine-d-amphetamine or a pharmaceutically acceptable
salt thereof for the treatment of ADHD; (b) sensitizing a doctor to
a product claim that the oral pharmaceutical composition has a
lower interpatient variation than an oral formulation containing
d-amphetamine; (c) diagnosing a patient as suffering from ADHD by
the doctor; (d) selectively prescribing the oral pharmaceutical
composition to the patient by the sensitized doctor in view of the
diagnosis of the patient; and (e) orally administering the
prescribed oral pharmaceutical composition to the patient.
[0021] Yet another embodiment is a method for treating a patient
suffering from ADHD comprising the steps of:
(a) purchasing an oral pharmaceutical composition comprising
L-lysine-d-amphetamine or a pharmaceutically acceptable salt
thereof; (b) tendering a prescription for the patient suffering
from ADHD before completing the purchase transaction, the
prescription having been received in response to a diagnosis of
ADHD by a doctor, the doctor (1) having been sensitized to a
product claim that the oral pharmaceutical composition has a lower
interpatient variation than an oral formulation containing
d-amphetamine, and (2) having selectively prescribed the oral
pharmaceutical composition; and (c) orally administering the
purchased oral pharmaceutical composition to the patient.
BRIEF DESCRIPTION OF DRAWINGS
[0022] FIG. 1. Synthesis of peptide amphetamine conjugates.
[0023] FIG. 2. Synthesis of lysine amphetamine dimesylate.
[0024] FIG. 3. Synthesis of lysine amphetamine HCl.
[0025] FIG. 4. Synthesis of serine amphetamine conjugate.
[0026] FIG. 5. Synthesis of phenylalanine amphetamine
conjugate.
[0027] FIG. 6. Synthesis of triglycine amphetamine conjugate.
[0028] FIG. 7. Plasma concentrations of d-amphetamine from
individual rats orally administered d-amphetamine or
L-lysine-d-amphetamine hydrochloride.
[0029] The following Figures (FIG. 8-FIG. 16) depict results
obtained from studies of oral administration of d-amphetamine
sulfate or L-lysine-d-amphetamine dimesylate to rats (ELISA
analysis):
[0030] FIG. 8. Plasma concentrations of d-amphetamine (at dose 1.5
mg/kg d-amphetamine base).
[0031] FIG. 9. Plasma concentrations of d-amphetamine (at dose 3
mg/kg d-amphetamine base).
[0032] FIG. 10. Plasma concentrations of d-amphetamine (at dose 6
mg/kg d-amphetamine base).
[0033] FIG. 11. Plasma concentrations of d-amphetamine (at dose 12
mg/kg d-amphetamine base).
[0034] FIG. 12. Plasma concentrations of d-amphetamine (at dose 30
mg/kg d-amphetamine base).
[0035] FIG. 13. Plasma concentrations of d-amphetamine (at dose 60
mg/kg d-amphetamine base).
[0036] FIG. 14. Percent bioavailability (AUC and C.sub.max) of
L-lysine-d-amphetamine dimesylate compared to d-amphetamine sulfate
at doses 1.5, 3, 6, 12, 30, and 60 mg/kg d-amphetamine base.
[0037] FIG. 15. Plasma concentrations of d-amphetamine at
30-minutes post-dose for escalating doses of d-amphetamine
base.
[0038] FIG. 16. Plasma concentrations of d-amphetamine (at dose 60
mg/kg d-amphetamine base).
[0039] FIG. 17. Plasma concentrations of d-amphetamine following
intranasal administration of L-lysine-d-amphetamine hydrochloride
or d-amphetamine sulfate (at dose 3 mg/kg d-amphetamine base) to
rats (ELISA analysis).
[0040] FIG. 18. Plasma concentrations of d-amphetamine following
intranasal administration of L-lysine-d-amphetamine dimesylate or
d-amphetamine sulfate (at dose 3 mg/kg d-amphetamine base) to rats
(ELISA analysis).
[0041] FIG. 19. Plasma concentrations of d-amphetamine following
bolus intravenous administration of L-lysine-d-amphetamine
dimesylate or d-amphetamine sulfate (at dose 1.5 mg/kg
d-amphetamine base) to rats (ELISA analysis).
[0042] FIG. 20. Plasma concentrations of d-amphetamine levels
following oral administration of Dexedrine Spansule.RTM. capsules,
crushed Dexedrine Spansule.RTM. capsules, or L-lysine-d-amphetamine
dimesylate (at dose 3 mg/kg d-amphetamine base) to rats (ELISA
analysis).
[0043] The following Figures (FIG. 21-FIG. 30) depict results
obtained from studies of oral administration of d-amphetamine
sulfate or L-lysine-d-amphetamine dimesylate to rats (LC/MS/MS
analysis):
[0044] FIG. 21A and FIG. 21B. Plasma concentrations of
d-amphetamine in ng/mL (FIG. 21A) and in nM (FIG. 21B) (at dose 1.5
mg/kg d-amphetamine base).
[0045] FIG. 22A and FIG. 22B. Plasma concentrations of
d-amphetamine in ng/mL (FIG. 22A) and in nM (FIG. 22B) (at dose 3
mg/kg d-amphetamine base).
[0046] FIG. 23A and FIG. 23B. Plasma concentrations of
d-amphetamine in ng/mL (FIG. 23A) and in nM (FIG. 23B) (at dose 6
mg/kg d-amphetamine base).
[0047] FIG. 24A and FIG. 24B. Plasma concentrations of
d-amphetamine in ng/mL (FIG. 24A) and in nM (FIG. 24B) (at dose 12
mg/kg d-amphetamine base).
[0048] FIG. 25A and FIG. 25B. Plasma concentrations of
d-amphetamine in ng/mL (FIG. 25A) and in nM (FIG. 25B) (at dose 60
mg/kg d-amphetamine base).
[0049] FIG. 26. Comparative bioavailability (C.sub.max) of
L-lysine-d-amphetamine and d-amphetamine in proportion to
escalating human equivalent doses.
[0050] FIG. 27. Comparative bioavailability (AUC.sub.inf) of
L-lysine-d-amphetamine and d-amphetamine in proportion to
escalating doses of d-amphetamine base.
[0051] FIG. 28. Comparative bioavailability (AUC.sub.inf) of
L-lysine-d-amphetamine and d-amphetamine in proportion to
escalating human equivalent doses.
[0052] FIG. 29. Comparative bioavailability (C.sub.max) of intact
L-lysine-d-amphetamine in proportion to escalating human equivalent
doses.
[0053] FIG. 30. Comparative bioavailability (AUC.sub.inf) of intact
L-lysine-d-amphetamine in proportion to escalating human equivalent
doses.
[0054] FIG. 31. Plasma concentrations of d-amphetamine following
intranasal administration of L-lysine-d-amphetamine dimesylate or
d-amphetamine sulfate (at dose 3 mg/kg d-amphetamine base) to rats
(LC/MS/MS analysis).
[0055] FIG. 32A and FIG. 32B. Plasma concentrations of
d-amphetamine and L-lysine-d-amphetamine in ng/mL (FIG. 32A) and in
nM (FIG. 32B), following intranasal administration of
L-lysine-d-amphetamine dimesylate or d-amphetamine sulfate (at dose
3 mg/kg d-amphetamine base) to rats (LC/MS/MS analysis).
[0056] FIG. 33. Plasma concentrations of d-amphetamine following
bolus intravenous administration of L-lysine-d-amphetamine
dimesylate or d-amphetamine sulfate (at dose 1.5 mg/kg
d-amphetamine base) to rats (LC/MS/MS analysis).
[0057] FIG. 34A and FIG. 34B. Plasma concentrations of
d-amphetamine in ng/mL (FIG. 34A) and in nM (FIG. 34B), following
intravenous administration of L-lysine-d-amphetamine dimesylate or
d-amphetamine sulfate (at dose 1.5 mg/kg d-amphetamine base) to
rats (LC/MS/MS analysis).
[0058] The following Figures (FIG. 35-FIG. 40) depict results
obtained from studies of oral and intravenous administration (at
dose 1 mg/kg d-amphetamine base) of d-amphetamine sulfate or
L-lysine-d-amphetamine dimesylate to conscious male beagle dogs
(LC/MS/MS analysis):
[0059] FIG. 35. Mean plasma concentration time profile of
L-lysine-d-amphetamine following intravenous or oral administration
of L-lysine-d-amphetamine (n=3).
[0060] FIG. 36. Plasma concentration time profile of d-amphetamine
following intravenous or oral administration of
L-lysine-d-amphetamine (n=3).
[0061] FIG. 37A and FIG. 37B. Mean plasma concentration time
profile of L-lysine-d-amphetamine and d-amphetamine levels in ng/ml
(FIG. 37A) and in nM (FIG. 37B), following intravenous
administration of L-lysine-d-amphetamine (n=3).
[0062] FIG. 38A and FIG. 38B. Mean plasma concentration time
profile of L-lysine-d-amphetamine and d-amphetamine levels in ng/ml
(FIG. 38A) and in nM (FIG. 38B), following oral administration of
L-lysine-d-amphetamine (n=3).
[0063] FIG. 39A and FIG. 39B. Individual plasma concentration time
profile of L-lysine-d-amphetamine following intravenous
administration (FIG. 39A) or oral administration (FIG. 39B) of
L-lysine-d-amphetamine.
[0064] FIG. 40A and FIG. 40B. Individual plasma concentration time
profile of d-amphetamine following intravenous administration (FIG.
40A) or oral administration (FIG. 40B) of
L-lysine-d-amphetamine.
[0065] FIG. 41. Plasma concentrations of d-amphetamine following
oral administration of L-lysine-d-amphetamine dimesylate or
d-amphetamine sulfate (at dose 1.8 mg/kg d-amphetamine base) to
male dogs.
[0066] FIG. 42. Plasma concentrations of d-amphetamine following
oral administration of L-lysine-d-amphetamine dimesylate or
d-amphetamine sulfate (at dose 1.8 mg/kg d-amphetamine base) to
female dogs.
[0067] FIG. 43. Mean blood pressure following intravenous injection
of increasing amounts of L-lysine-d-amphetamine dimesylate or
d-amphetamine in male and female dogs.
[0068] FIG. 44. Left ventricular blood pressure following
intravenous injection of increasing amounts of
L-lysine-d-amphetamine dimesylate or d-amphetamine in male and
female dogs.
[0069] The following Figures (FIG. 45-FIG. 49) depict results
obtained from studies of oral (at dose 6 mg/kg d-amphetamine base),
intranasal (at dose 1 mg/kg d-amphetamine base), and intravenous
administration (at dose 1 mg/kg d-amphetamine base) of
d-amphetamine sulfate or L-lysine-d-amphetamine hydrochloride to
rats:
[0070] FIG. 45. Locomotor activity of rats following oral
administration (5 hour time-course).
[0071] FIG. 46. Locomotor activity of rats following oral
administration (12 hour time-course).
[0072] FIG. 47. Locomotor activity of rats following intranasal
administration (1 hour time-course).
[0073] FIG. 48. Locomotor activity of rats following intranasal
administration (with carboxymethylcellulose) (2 hour
time-course).
[0074] FIG. 49. Locomotor activity of rats following intravenous
administration (3 hour time-course).
[0075] The following Figures (FIG. 50-FIG. 58) depict results
obtained from studies of oral, intranasal, and intravenous
administration of d-amphetamine or amphetamine conjugate
hydrochloride salts to rats (ELISA analysis):
[0076] FIG. 50. Intranasal bioavailability of abuse-resistant
amphetamine amino acid, di-, and tri-peptide conjugates.
[0077] FIG. 51. Oral bioavailability of abuse-resistant amphetamine
amino acid, di-, and tri-peptide conjugates.
[0078] FIG. 52. Intravenous bioavailability of an abuse-resistant
amphetamine tri-peptide conjugate.
[0079] FIG. 53. Intranasal bioavailability of an abuse-resistant
amphetamine amino acid conjugate.
[0080] FIG. 54. Oral bioavailability of an abuse-resistant
amphetamine amino acid conjugate.
[0081] FIG. 55. Intravenous bioavailability of an abuse-resistant
amphetamine amino acid conjugate.
[0082] FIG. 56. Intranasal bioavailability of an abuse-resistant
amphetamine amino tri-peptide conjugate.
[0083] FIG. 57. Intranasal bioavailability of abuse-resistant
amphetamine amino acid-, and di-peptide conjugates.
[0084] FIG. 58. Intranasal bioavailability of an abuse-resistant
amphetamine di-peptide conjugate containing D- and L-amino acid
isomers.
[0085] FIG. 59A and FIG. 59B. Plasma concentrations of
d-amphetamine and L-lysine-d-amphetamine in ng/mL for the serum
levels (FIG. 59A) and in ng/g for brain tissue (FIG. 59B),
following oral administration of L-lysine-d-amphetamine
hydrochloride or d-amphetamine sulfate (at dose 5 mg/kg
d-amphetamine base) to rats (LC/MS/MS analysis).
[0086] FIG. 60. Steady-state plasma d-amphetamine and
L-lysine-d-amphetamine levels obtained from clinical studies of
oral administration of L-lysine-d-amphetamine dimesylate 70 mg to
humans (LC/MS/MS analysis).
[0087] The following Figures (FIG. 61-FIG. 70) depict results
obtained from clinical studies of oral administration of
L-lysine-d-amphetamine dimesylate to humans (LC/MS/MS
analysis):
[0088] FIG. 61A and FIG. 61B. Plasma d-amphetamine and
L-lysine-d-amphetamine levels (FIG. 61A, ng/mL; FIG. 61B, nM) over
a 72 hour period following oral administration of
L-lysine-d-amphetamine (25 mg L-lysine-d-amphetamine dimesylate
containing 7.37 mg d-amphetamine base) to humans.
[0089] FIG. 62A and FIG. 62B. Plasma d-amphetamine and
L-lysine-d-amphetamine levels (FIG. 62A, ng/mL; FIG. 62B, nM) over
a 72 hour period following oral administration of
L-lysine-d-amphetamine (75 mg L-lysine-d-amphetamine dimesylate
containing 22.1 mg d-amphetamine base) to humans.
[0090] FIG. 63A and FIG. 63B. Plasma d-amphetamine levels (FIG.
63A, 0-12 hours; FIG. 63B, 0-72 hours) following oral
administration of L-lysine-d-amphetamine (75 mg
L-lysine-d-amphetamine dimesylate containing 22.1 mg d-amphetamine
base) or Adderall XR.RTM. (35 mg containing 21.9 mg amphetamine
base) to humans.
[0091] FIG. 64A and FIG. 64B. Plasma d-amphetamine levels (FIG.
64A, 0-12 hours; FIG. 64B, 0-72 hours) following oral
administration of L-lysine-d-amphetamine (75 mg
L-lysine-d-amphetamine dimesylate containing 22.1 mg d-amphetamine
base) or Dexedrine Spansule.RTM. (30 mg containing 22.1 mg
amphetamine base) to humans.
[0092] FIG. 65. Mean plasma concentration of d-amphetamine after
oral administration of single 30 mg, 50 mg, and 70 mg doses of
L-lysine-d-amphetamine dimesylate under fasted conditions to
pediatric patients with ADHD.
[0093] FIG. 66. Relationship between the dose-normalized AUC of
d-amphetamine and gender after oral administration of
L-lysine-d-amphetamine dimesylate capsules once daily to healthy
adult volunteers and children with ADHD.
[0094] FIG. 67. Relationship between the dose-normalized maximum
plasma concentration of d-amphetamine and gender after oral
administration of L-lysine-d-amphetamine dimesylate capsules once
daily to healthy adult volunteers and children with ADHD.
[0095] FIG. 68. Relationship between the dose-normalized time to
maximum concentration of d-amphetamine and gender after oral
administration of L-lysine-d-amphetamine dimesylate capsules once
daily to healthy adult volunteers and children with ADHD.
[0096] FIG. 69. ADHD-RS at endpoint for pediatric clinical
study.
[0097] FIG. 70. SKAMP score (efficacy) vs. time for pediatric
clinical study.
[0098] FIGS. 71 and 72. Optical micrographs and scanning electron
micrographs, respectively, of L-lysine-d-amphetamine dimesylate
prepared according to the procedure described in Example 38.
[0099] FIGS. 73 and 74. Optical micrographs of
L-lysine-d-amphetamine dimesylate prepared according to the
procedure described in Example 39.
[0100] FIGS. 75 and 76. Scanning electron micrographs of
L-lysine-d-amphetamine dimesylate prepared according to the
procedure described in Example 39.
[0101] FIG. 77. X-ray powder diffractogram (XRPD) of
L-lysine-d-amphetamine dimesylate prepared according to the
procedure described in Example 39.
[0102] FIG. 78. Differential scanning calorimetry (DSC) thermogram
of lisdexamphetamine dimesylate prepared according to the procedure
described in Example 38
[0103] FIGS. 79 and 80. Fourier transform (FT)-Raman Spectra, at
high and low sensitivity, of lisdexamphetamine dimesylate prepared
according to the procedure described in Example 39.
[0104] FIG. 81. ORTEP drawing of the contents of the asymmetric
unit of the lisdexamphetamine dimesylate crystal structure. Atoms
are represented by 50% probability anisotropic thermal
ellipsoids.
[0105] FIG. 82. ORTEP drawing of the two lisdexamphetamine
molecules observed in the crystal structure. Atoms are represented
by 50% probability anisotropic thermal ellipsoids.
[0106] FIG. 83. Packing diagram of lisdexamphetamine dimesylate
viewed down the crystallographic a axis.
[0107] FIG. 84. Packing diagram of lisdexamphetamine dimesylate
viewed down the crystallographic b axis.
[0108] FIG. 85. Packing diagram of lisdexamphetamine dimesylate
viewed down the crystallographic c axis.
[0109] FIG. 86. Comparison of calculated XRPD spectrum from single
crystal data (top) to experimental XRPD spectrum of
lisdexamphetamine dimesylate prepared according to example 39.
[0110] FIG. 87. Synthesis of lysine amphetamine dimesylate.
[0111] FIG. 88 is a graph showing the mean change in SKAMP
deportment score from the pre-dose measurement over 13 hours after
dosing, for both placebo and L-lysine-d-amphetamine dimesylate.
[0112] FIG. 89 is a graph showing the mean change in SKAMP
attention score from the pre-dose measurement over 13 hours after
dosing, for both placebo and L-lysine-d-amphetamine dimesylate.
[0113] FIG. 90 is a graph showing the mean change in PERMP-A scores
from the pre-dose measurement over 13 hours after dosing, for both
placebo and L-lysine-d-amphetamine dimesylate.
[0114] FIG. 91 is a graph showing the mean change in PERMP-C scores
from the pre-dose measurement over 13 hours after dosing, for both
placebo and L-lysine-d-amphetamine dimesylate.
[0115] FIG. 92 is a histogram analysis of CGI-I scores for a
cross-over treatment phase (between lysine-d-amphetamine dimesylate
and placebo).
[0116] FIG. 93 is a graph showing the mean plasma d-amphetamine
concentration over time, for individual subjects.
DETAILED DESCRIPTION OF THE INVENTION
[0117] The invention provides amphetamine prodrugs comprising
amphetamine covalently bound to a chemical moiety. The amphetamine
prodrugs can also be characterized as conjugates in that they
possess a covalent attachment. They may also be characterized as
conditionally bioreversible derivatives ("CBDs") in that the
amphetamine prodrug preferably remains inactive until oral
administration releases the amphetamine from the chemical
moiety.
[0118] In one embodiment, the invention provides an amphetamine
prodrug of Formula I:
A-X.sub.n-Z.sub.m (I)
[0119] wherein A is an amphetamine;
[0120] each X is independently a chemical moiety;
[0121] each Z is independently a chemical moiety that acts as an
adjuvant and is different from at least one X;
[0122] n is an increment from 1 to 50, preferably 1 to 10; and
[0123] m is an increment from 0 to 50, preferably 0.
[0124] When m is 0, the amphetamine prodrug is a compound of
Formula (II):
A-X.sub.n (II)
[0125] wherein each X is independently a chemical moiety.
[0126] Formula (II) can also be written to designate the chemical
moiety that is physically attached to the amphetamine:
A-X.sub.1-(X).sub.n-1 (III)
[0127] wherein A is an amphetamine; X.sub.1 is a chemical moiety,
preferably a single amino acid; each X is independently a chemical
moiety that is the same as or different from X.sub.1; and n is an
increment from 1 to 50.
[0128] The amphetamine, A, can be any of the sympathomimetic
phenethylamine derivatives which have central nervous system
stimulant activity such as amphetamine, or any derivative, analog,
or salt thereof. Exemplary amphetamines include, but are not
limited to, amphetamine, methamphetamine, methylphenidate,
p-methoxyamphetamine, methylenedioxyamphetamine,
2,5-dimethoxy-4-methylamphetamine, 2,4,5-trimethoxyamphetamine, and
3,4-methylenedioxymethamphetamine, N-ethylamphetamine,
fenethylline, benzphetamine, and chlorphentermine as well as the
amphetamine compounds of Adderall.RTM.; actedron; actemin; adipan;
akedron; allodene; alpha-methyl-(.+-.)-benzeneethanamine;
alpha-methylbenzeneethanamine; alpha-methylphenethylamine;
amphetamine; amphate; anorexine; benzebar; benzedrine; benzyl
methyl carbinamine; benzolone; beta-amino propylbenzene;
beta-phenylisopropylamine; biphetamine; desoxynorephedrine;
dietamine; DL-amphetamine; elastonon; fenopromin; finam; isoamyne;
isomyn; mecodrin; monophos; mydrial; norephedrane; novydrine;
obesin; obesine; obetrol; octedrine; oktedrin; phenamine;
phenedrine; phenethylamine, alpha-methyl-; percomon; profamina;
profetamine; propisamine; racephen; raphetamine; rhinalator,
sympamine; simpatedrin; simpatina; sympatedrine; and weckamine.
Preferred amphetamines include methamphetamine, methylphenidate,
and amphetamine, with amphetamine being most preferred.
##STR00001##
[0129] The amphetamine can have any stereogenic configuration,
including both dextro- and levo-isomers. The dextro-isomer,
particularly dextroamphetamine, is preferred.
[0130] Preferably, the amphetamine is an amphetamine salt.
Pharmaceutically acceptable salts, e.g., non-toxic, inorganic and
organic acid addition salts, are known in the art. Exemplary salts
include, but are not limited to, 2-hydroxyethanesulfonate,
2-naphthalenesulfonate, 3-hydroxy-2-naphthoate, 3-phenylpropionate,
acetate, adipate, alginate, amsonate, aspartate, benzenesulfonate,
benzoate, besylate, bicarbonate, bisulfate, bitartrate, borate,
butyrate, calcium edetate, camphorate, camphorsulfonate, camsylate,
carbonate, citrate, clavulariate, cyclopentanepropionate,
digluconate, dodecylsulfate, edetate, edisylate, estolate, esylate,
ethanesulfonate, finnarate, gluceptate, glucoheptanoate, gluconate,
glutamate, glycerophosphate, glycollylarsanilate, hemisulfate,
heptanoate, hexafluorophosphate, hexanoate, hexylresorcinate,
hydrabamine, hydrobromide, hydrochloride, hydroiodide,
hydroxynaphthoate, iodide, isothionate, lactate, lactobionate,
laurate, laurylsulphonate, malate, maleate, mandelate, mesylate,
methanesulfonate, methylbromide, methylnitrate, methylsulfate,
mucate, naphthylate, napsylate, nicotinate, nitrate,
N-methylglucamine ammonium salt, oleate, oxalate, palmitate,
pamoate, pantothenate, pectinate, persulfate, phosphate,
phosphateldiphosphate, picrate, pivalate, polygalacturonate,
propionate, p-toluenesulfonate, saccharate, salicylate, stearate,
subacetate, succinate, sulfate, sulfosaliculate, suramate, tannate,
tartrate, teoclate, thiocyanate, tosylate, triethiodide,
undecanoate, and valerate salts, and the like. (See Berge et al.
(1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19). A preferred
amphetamine salt is the mesylate salt (e.g., as in
L-lysine-d-amphetamine dimesylate).
[0131] Particular salts may be less hygroscopic thereby
facilitating handling. In a preferred embodiment, the amphetamine
prodrug has a water content (Karl Fischer analysis) of about 0% to
about 5%, about 0.1% to about 3%, about 0.25% to about 2%, or
increments therein. When the amphetamine prodrug is formulated into
a pharmaceutical composition, the pharmaceutical composition
preferably has a water content of about 1% to about 10%, about 1%
to about 8%, about 2% to about 7%, or increments therein.
DEFINITIONS
[0132] The term "crystal" refers to a form of a solid state of
matter, which is distinct from its amorphous solid state. Crystals
display characteristic features including a lattice structure,
characteristic shapes and optical properties such as refractive
index. A crystal consists of atoms arranged in a pattern that
repeats periodically in three dimensions.
[0133] An "anhydrous crystal form" lacks bound water molecules.
[0134] The term "polymorph" refers to crystallographically distinct
forms of a substance.
[0135] The term "amine" refers to a --NH.sub.2 group.
[0136] The D.sub.10, D.sub.50 and D.sub.90 represent the 10.sup.th
percentile, median or the 50th percentile and the 90th percentile
of the particle size distribution, respectively, as measured by
diameter. That is, the D.sub.10 (D.sub.50, D.sub.90) is a value on
the distribution such that 10% (50%, 90%) of the particles have a
volume of this value or less.
[0137] An "effective amount of drug" is an amount of
lisdexamphetamine dimesylate which is effective to treat or prevent
a condition in a living organism to whom it is administered over
some period of time, e.g., provides a therapeutic effect during a
desired dosing interval.
[0138] The term "treating" includes: (1) preventing or delaying the
appearance of clinical symptoms of the state, disorder or condition
developing in an animal that may be afflicted with or predisposed
to the state, disorder or condition but does not yet experience or
display clinical or subclinical symptoms of the state, disorder or
condition; (2) inhibiting the state, disorder or condition (i.e.,
arresting, reducing or delaying the development of the disease, or
a relapse thereof in case of maintenance treatment, of at least one
clinical or subclinical symptom thereof); and/or (3) relieving the
condition (i.e., causing regression of the state, disorder or
condition or at least one of its clinical or subclinical symptoms).
The benefit to a patient to be treated is either statistically
significant or at least perceptible to the patient or to the
physician.
[0139] As used herein in connection with a measured quantity, the
term "about" refers to the normal variation in that measured
quantity that would be expected by the skilled artisan making the
measurement and exercising a level of care commensurate with the
objective of the measurement and the precision of the measuring
equipment used. Unless otherwise indicated, "about" refers to a
variation of .+-.10% of the value provided.
[0140] Throughout this application, the term "increment" is used to
define a numerical value in varying degrees of precision, e.g., to
the nearest 10, 1, 0.1, 0.01, etc. The increment can be rounded to
any measurable degree of precision. For example, the range 1 to 100
or increments therein includes ranges such as 20 to 80, 5 to 50,
0.4 to 98, and 0.04 to 98.05.
[0141] The amphetamine is bound to one or more chemical moieties,
denominated X and Z. A chemical moiety can be any moiety that
decreases the pharmacological activity of amphetamine while bound
to the chemical moiety as compared to unbound (free) amphetamine.
The attached chemical moiety can be either naturally occurring or
synthetic. Exemplary chemical moieties include, but are not limited
to, peptides, including single amino acids, dipeptides,
tripeptides, oligopeptides, and polypeptides; glycopeptides;
carbohydrates; lipids; nucleosides; nucleic acids; and vitamins.
Preferably, the chemical moiety is generally recognized as safe
("GRAS").
[0142] "An extended release mixed amphetamine composition," such as
Adderall.TM. XR, is a composition that contains at least 2
amphetamine salts. The composition allows for extended release of
amphetamine into the bloodstream.
[0143] "Carbohydrates" include sugars, starches, cellulose, and
related compounds, e.g., (CH.sub.2O).sub.n wherein n is an integer
larger than 2, and C.sub.n(H.sub.2O).sub.n-1 wherein n is an
integer larger than 5. The carbohydrate can be a monosaccharide,
disaccharide, oligosaccharide, polysaccharide, or a derivative
thereof (e.g., sulfo- or phospho-substituted). Exemplary
carbohydrates include, but are not limited to, fructose, glucose,
lactose, maltose, sucrose, glyceraldehyde, dihydroxyacetone,
erythrose, ribose, ribulose, xylulose, galactose, mannose,
sedoheptulose, neuraminic acid, dextrin, and glycogen.
[0144] A "glycopeptide" is a carbohydrate linked to an
oligopeptide. Similarly, the chemical moiety can also be a
glycoprotein, glyco-amino-acid, or glycosyl-amino-acid. A
"glycoprotein" is a carbohydrate (e.g., a glycan) covalently linked
to a protein. A "glyco-amino-acid" is a carbohydrate (e.g., a
saccharide) covalently linked to a single amino acid. A
"glycosyl-amino-acid" is a carbohydrate (e.g., a saccharide) linked
through a glycosyl linkage (O--, N--, or S--) to an amino acid.
[0145] A "peptide" includes a single amino acid, a dipeptide, a
tripeptide, an oligopeptide, a polypeptide, or a carrier peptide.
An oligopeptide includes from 2 to 70 amino acids.
[0146] The term "amino acid" includes both naturally occurring
(i.e., the 20 amino acids used for protein synthesis) and
nonnaturally occurring amino acids, synthetically produced amino
acids.
[0147] Preferably, the chemical moiety is a peptide, more
particularly a single amino acid, a dipeptide, or a tripeptide. The
peptide preferably comprises fewer than 70 amino acids, fewer than
50 amino acids, fewer than 10 amino acids, or fewer than 4 amino
acids. When the chemical moiety is one or more amino acids, the
amphetamine is preferably bound to lysine, serine, phenylalanine,
or glycine. In another embodiment, the amphetamine is preferably
bound to lysine, glutamic acid, or leucine. In one embodiment, the
amphetamine is bound to lysine and optional additional chemical
moieties, e.g., additional amino acids. In a preferred embodiment,
the amphetamine is bound to a single lysine amino acid.
[0148] In one embodiment, the chemical moiety is from 1 to 12 amino
acids, preferably 1 to 8 amino acids. In another embodiment, the
number of amino acids is 1, 2, 3, 4, 5, 6, or 7. In another
embodiment, the molecular weight of the chemical moiety is below
about 2,500 kD, more preferably below about 1,000 kD, and most
preferably below about 500 kD.
[0149] Each amino acid can be any one of the L- or D-enantiomers,
preferably L-enantiomers, of the naturally occurring amino acids:
alanine (Ala or A), arginine (Arg or R), asparagine (Asn or N),
aspartic acid (Asp or D), cysteine (Cys or C), glycine (Gly or G),
glutamic acid (Glu or E), glutamine (Gln or Q), histidine (His or
H), isoleucine (Ile or I), leucine (Leu or L), lysine (Lys or K),
methionine (Met or M), proline (Pro or P), phenylalanine (Phe or
F), serine (Ser or S), tryptophan (Trp or W), threonine (Thr or T),
tyrosine (Tyr or Y), and valine (Val or V). In a preferred
embodiment, the peptide comprises only naturally occurring amino
acids and/or only L-amino acids. Each amino acid can be an
unnatural, non-standard, or synthetic amino acids, such as
aminohexanoic acid, biphenylalanine, cyclohexylalanine,
cyclohexylglycine, diethylglycine, dipropylglycine,
2,3-diaminoproprionic acid, homophenylalanine, homoserine,
homotyrosine, naphthylalanine, norleucine, ornithine, phenylalanine
(4-fluoro), phenylalanine(2,3,4,5,6-pentafluoro),
phenylalanine(4-nitro), phenylglycine, pipecolic acid, sarcosine,
tetrahydroisoquinoline-3-carboxylic acid, and tert-leucine.
Preferably, synthetic amino acids with alkyl side chains are
selected from C.sub.1-C.sub.17 alkyls, preferably C.sub.1-C.sub.6
alkyls. In one embodiment, the peptide comprises one or more amino
acid alcohols, e.g., serine and threonine. In another embodiment,
the peptide comprises one or more N-methyl amino acids, e.g.,
N-methyl aspartic acid.
[0150] In one embodiment, the peptides are utilized as base short
chain amino acid sequences and additional amino acids are added to
the terminus or side chain. In another embodiment, the peptide may
have an one or more amino acid substitutions. Preferably, the
substitute amino acid is similar in structure, charge, or polarity
to the replaced amino acid. For instance, isoleucine is similar to
leucine, tyrosine is similar to phenylalanine, serine is similar to
threonine, cysteine is similar to methionine, alanine is similar to
valine, lysine is similar to arginine, asparagine is similar to
glutamine, aspartic acid is similar to glutamic acid, histidine is
similar to proline, and glycine is similar to tryptophan.
[0151] The peptide can comprise a homopolymer or heteropolymer of
naturally occurring or synthetic amino acids. For example, the side
chain attachment of amphetamine to the peptide can be a homopolymer
or heteropolymer containing glutamic acid, aspartic acid, serine,
lysine, cysteine, threonine, asparagine, arginine, tyrosine, or
glutamine.
[0152] Exemplary peptides include Lys, Ser, Phe, Gly-Gly-Gly,
Leu-Ser, Leu-Glu, homopolymers of Glu and Leu, and heteropolymers
of (Glu).sub.n-Leu-Ser. In a preferred embodiment, the peptide is
Lys, Ser, Phe, or Gly-Gly-Gly.
[0153] In one embodiment, the chemical moiety has one or more free
carboxy and/or amine terminal and/or side chain group other than
the point of attachment to the amphetamine. The chemical moiety can
be in such a free state, or an ester or salt thereof.
[0154] The chemical moiety can be covalently attached to the
amphetamine either directly or indirectly through a linker.
Covalent attachment may comprise an ester or carbonate bond. The
site of attachment typically is determined by the functional
group(s) available on the amphetamine. For example, a peptide can
be attached to an amphetamine via the N-terminus, C-terminus, or
side chain of an amino acid. For additional methods of attaching
amphetamine to various exemplary chemical moieties, see U.S.
application Ser. No. 10/156,527, PCT/US03/05524, and
PCT/US03/05525, each of which is hereby incorporated by reference
in its entirety.
[0155] Synthesis of Lisdexamphetamine and Salts Thereof.
[0156] Lisdexamphetamine and salts thereof can be prepared from
L-lysine or a salt thereof as follows. The amine groups on the
L-lysine or a salt thereof are protected, for example, by reaction
with di-tert-butyl dicarbonate. Suitable amine protecting groups
include, but are not limited to, Boc (--C(O)OC(CH.sub.3).sub.3),
Boc (tert-butyloxycarbonyl, --C(O)OC(CH.sub.3).sub.3), Cbz
(benzyloxycarbonyl, --C(O)OCH.sub.2Ph), Alloc (allyloxycarbonyl,
--C(O)OCH.sub.2CH.dbd.CH.sub.2), Fmoc (Fluorenylmethyloxycarbonyl,
--C(O)OCH.sub.2-fluorene) or Teoc (Trimethylsilylethyl-oxycarbonyl,
--C(O)OCH.sub.2CH.sub.2Si(CH.sub.3).sub.3). In addition, amine
protecting groups given in J. Chem. Soc., Perkin Trans. Vol. 1,
1998, p. 4005-4037, hereby incorporated by reference in its
entirety. Preferably this reaction is performed in the presence of
a base, such as sodium hydroxide.
[0157] The di-amine protected L-lysine is then subjected to an acid
activation followed by an amidation reaction with d-amphetamine to
form a di-amine protected lisdexamphetamine. The amidation reaction
can be performed either by first activating the acid group of the
protected L-lysine, and then amidating the activated acid by, for
example, reacting it with d-amphetamine ("two-step procedure"), or
in a "one-pot procedure" in which the protected L-lysine,
d-amphetamine, coupling reagent, and optional additive are all
combined simultaneously. For the two-step procedure, the acid group
of the protected L-lysine can be activated, for example, by
reaction with DCC (dicyclohexylcarbodiimide), CDI
(carbonyldiimidazole), EDCI
(1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide), PyBOP
(benzotriazol-1-yloxytris(pyrrolidino)phosphonium
hexafluorophosphate), or HBTU
(O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate) in addition to, for example,
N-hydroxysuccinimide, N-hydroxybenzotriazole, or a phenol such as
p-nitrophenol, or by reaction with, for instance,
iso-butylchloroformate. Preferably, the activated ester is then
isolated before being allowed to react with d-amphetamine.
[0158] In the one-pot procedure, the activation of the protected
L-lysine acid and subsequent amidation can be performed by mixing
the protected L-lysine, d-amphetamine, coupling reagent and
optional additive simultaneously. The coupling agent may be, for
example, DCC (dicyclohexylcarbodiimide), CDI (carbonyldiimidazole),
EDCI (1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide), PyBOP
(benzotriazol-1-yloxytris(pyrrolidino)phosphonium
hexafluorophosphate), or HBTU
(O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate). The optional additive may be, for example,
N-hydroxysuccinimide, N-hydroxybenzotriazole, or a phenol such as
p-nitrophenol. Preferably, the acid activation step is performed in
the presence of a dehydration agent, such as
dicyclohexylcarbodiimide (DCC). In either the two-step or one
one-pot method, the amidating step can be performed in the presence
of an organic base, such as N-methyl-morpholine (NMM).
[0159] The di-amine protected lisdexamphetamine is then deprotected
and optionally converted to a salt. The deprotection and salt
conversion can be performed in a single reaction. For example, the
di-amine protected lisdexamphetamine can be reacted with methane
sulfonic acid to form lisdexamphetamine dimesylate. Preferably,
this reaction is performed in the presence of an alcohol, such as
isopropanol.
[0160] The amphetamine prodrug compounds described above can be
synthesized as described in Examples 1, 38 and 39 and FIG. 1 and
FIG. 87. Preferably, additional purification and/or crystallization
steps are not necessary to yield a highly pure product. In one
embodiment, the purity of the amphetamine prodrug is at least about
95%, more preferably at least about 96%, 97%, 98%, 98.5%, 99%,
99.5%, 99.9%, or increments therein. For the synthesis of
L-lysine-d-amphetamine, known impurities include
Lys-Lys-d-amphetamine, Lys(Lys)-d-amphetamine, d-amphetamine,
Lys(Boc)-d-amphetamine, Boc-Lys-d-amphetamine, and
Boc-Lys(Boc)-d-amphetamine. In one embodiment, the presence of any
single impurity is less than about 3%, more preferably less than
about 2%, 1%, 0.5%, 0.25%, 0.2%, 0.15%, 0.1%, 0.05%, or increments
therein (e.g., greater than 0% but less than 0.2, 0.1, or 0.05%).
The impurities and their structures are given in the table
below.
TABLE-US-00002 Common Name IUPAC Name Structure Lys(Lys)- Dex or
LysLysDex Isomer 1 (2S)-2,6-Diamino-N-{(5S)- 5-amino-6-[[(1S)-1-
methyl-2- phenylethyl]amino]-6- oxohexyl}hexanamide
trimethanesulfonate ##STR00002## LysLys-Dex or LysLysDex Isomer 2
(2S)-2,6-Diamino-N-{(1S)- 5-amino-1-[[((1S)-1- methyl-2-
phenylethyl)amino]carbon yl]pentyl} hexanamide trimethanesulfonate
##STR00003## BocLys-Dex or (monoBoc)L ys-Dex 1 tert-Butyl
(1S)-5-amino-1- {[((1S)-1-methyl-2- phenylethyl)amino]carbon
yl}pentylcarbamate methanesulfonate ##STR00004## Lys(Boc)- Dex or
(monoBoc)L ys-Dex 2 tert-Butyl (5S)-5-amino-6- {[(1S)-1-methyl-2-
phenylethyl]amino}-6- oxohexylcarbamate methanesulfonate
##STR00005## Boc- Lys(Boc)- Dex tert-Butyl (1S)-5-[(tert-
butoxycarbonyl)amino]-1- ({[(1S)-1-methyl-2-
phenylethyl]amino}carbonyl) pentylcarbamate ##STR00006##
Diastereomer (2R)-2,6-Diamino-N-[(1S)- 1-methyl-2-
phenylethyl]hexanamide dimethanesulfonate ##STR00007##
Dextroamphetamine methanesulfonate (1S)-1-Methyl-2-
phenylethylamine methanesulfonate ##STR00008##
[0161] In one embodiment, the amphetamine prodrug (a compound of
one of the formulas described above) may exhibit one or more of the
following advantages over free amphetamines. The amphetamine
prodrug may prevent overdose by exhibiting a reduced
pharmacological activity when administered at higher than
therapeutic doses, e.g., higher than the prescribed dose. Yet when
the amphetamine prodrug is administered at therapeutic doses, the
amphetamine prodrug may retain similar pharmacological activity to
that achieved by administering unbound amphetamine, e.g., Adderall
XR.RTM.. Also, the amphetamine prodrug may prevent abuse by
exhibiting stability under conditions likely to be employed by
illicit chemists attempting to release the amphetamine. The
amphetamine prodrug may prevent abuse by exhibiting reduced
bioavailability when it is administered via parenteral routes,
particularly the intravenous ("shooting"), intranasal ("snorting"),
and/or inhalation ("smoking") routes that are often employed in
illicit use. Thus, the amphetamine prodrug may reduce the euphoric
effect associated with amphetamine abuse. Thus, the amphetamine
prodrug may prevent and/or reduce the potential of abuse and/or
overdose when the amphetamine prodrug is used in a manner
inconsistent with the manufacturer's instructions, e.g., consuming
the amphetamine prodrug at a higher than therapeutic dose or via a
non-oral route of administration.
[0162] Use of phrases such as "decreased", "reduced", "diminished",
or "lowered" includes at least a 10% change in pharmacological
activity with greater percentage changes being preferred for
reduction in abuse potential and overdose potential. For instance,
the change may also be greater than 25%, 35%, 45%, 55%, 65%, 75%,
85%, 95%, 96%, 97%, 98%, 99%, or other increments greater than
10%.
[0163] The term "bioavailability" refers to the rate and extent to
which a drug is absorbed. One measurement of bioavailability is
defined by the fraction (F) of the dose that reaches systemic
circulation. Thus, in extreme cases, F=0 in drugs which are not
absorbed at all in the GI tract while for drugs that are completely
absorbed (and not metabolized by a first pass effect) F=1. The
bioavailability can also be calculated from the area under the
curve (AUC) of the serum level vs. time plot.
[0164] The coefficient of variation (CV) is typically used to
express the variability in bioavailability. This value is obtained
by expressing the standard deviation as a percentage of the
arithmetic mean.
[0165] Use of the phrase "similar pharmacological activity" means
that two compounds exhibit curves that have substantially the same
AUC, C.sub.max, T.sub.max, C.sub.min, and/or t.sub.1/2 parameters,
preferably within about 30% of each other, more preferably within
about 25%, 20%, 10%, 5%, 2%, 1%, or other increments less than
30%.
[0166] Preferably, the amphetamine prodrug exhibits an unbound
amphetamine oral bioavailability of at least about 60% AUC (area
under the curve), more preferably at least about 70%, 80%, 90%,
95%, 96%, 97%, 98%, 99%, or other increments greater than 60%.
Preferably, the amphetamine prodrug exhibits an unbound amphetamine
parenteral, e.g., intranasal, bioavailability of less than about
70% AUC, more preferably less than about 50%, 30%, 20%, 15%, 10%,
5%, 4%, 3%, 2%, 1%, or other increments less than 70%. For certain
treatments, it is desirable that the amphetamine prodrug exhibits
both the oral and parenteral bioavailability characteristics
described above. See, e.g., Table 61.
[0167] Preferably, the amphetamine prodrug remains inactive until
oral administration releases the amphetamine. Without being bound
by theory, it is believed that the amphetamine prodrug is inactive
because the attachment of the chemical moiety reduces binding
between the amphetamine and its biological target sites (e.g.,
human dopamine ("DAT") and norepinephrine ("NET") transporter
sites). (See Hoebel, B. G., L. Hernandez, et al., "Microdialysis
studies of brain norepinephrine, serotonin, and dopamine release
during ingestive behavior, Theoretical and clinical implications."
Ann NY Acad Sci 575: 171-91 (1989)). The chemical moiety attachment
may reduce binding between amphetamine and DAT and/or NET in part
because the amphetamine prodrug cannot cross the blood-brain
barrier. The amphetamine prodrug is activated by oral
administration, that is, the amphetamine is released from the
chemical moiety by hydrolysis, e.g., by enzymes in the stomach,
intestinal tract, or blood serum. Because oral administration
facilitates activation, activation is reduced when the amphetamine
prodrug is administered via parenteral routes often employed by
illegal users.
[0168] Further, it is believed that the amphetamine prodrug is
resistant to abuse and/or overdose due to a natural gating
mechanism at the site of hydrolysis, namely the gastrointestinal
tract. This gating mechanism is thought to allow the release of
therapeutic amounts of amphetamine from the amphetamine prodrug,
but limit the release of higher amounts of amphetamine.
[0169] In another embodiment, the toxicity of the amphetamine
prodrug is substantially lower than that of the unbound
amphetamine. For example, in a preferred embodiment, the acute
toxicity is 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold,
8-fold, 9-fold, 10-fold less, or increments therein less lethal
than oral administration of unbound amphetamine.
[0170] Preferably, the amphetamine prodrug provides a serum release
curve that does not increase above amphetamine's toxicity level
when administered at higher than therapeutic doses. The amphetamine
prodrug may exhibit a reduced rate of amphetamine absorption and/or
an increased rate of clearance compared to the free amphetamine.
The amphetamine prodrug may also exhibit a steady-state serum
release curve. Preferably, the amphetamine prodrug provides
bioavailability but prevents C.sub.max spiking or increased blood
serum concentrations. Pharmacokinetic parameters are described in
the Examples below, particularly the clinical pharmacokinetic
Examples. In one embodiment, the amphetamine prodrug provides
similar pharmacological activity to the clinically measured
pharmacokinetic activity of L-lysine-d-amphetamine dimesylate. For
example, the pharmacological parameters (AUC, C.sub.max, T.sub.max,
C.sub.min, and/or t.sub.1/2) are preferably within 80% to 125%, 80%
to 120%, 85% to 125%, 90% to 110%, or increments therein, of the
given values. It should be recognized that the ranges can, but need
not be symmetrical, e.g., 85% to 105%. For the pediatric study, the
pharmacokinetic parameters of d-amphetamine released from
L-lysine-d-amphetamine dimesylate are listed in Table 72.
[0171] The amphetamine prodrug may exhibit delayed and/or sustained
release characteristics. Delayed release prevents rapid onset of
pharmacological effects, and sustained release is a desirable
feature for particular dosing regimens, e.g., once a day regimens.
The amphetamine prodrug may achieve the release profile
independently. Alternatively, the amphetamine prodrug may be
pharmaceutically formulated to enhance or achieve such a release
profile. It may be desirable to reduce the amount of time until
onset of pharmacological effect, e.g., by formulation with an
immediate release product.
[0172] Accordingly, the invention also provides methods comprising
providing, administering, prescribing, or consuming an amphetamine
prodrug. The invention also provides pharmaceutical compositions
comprising an amphetamine prodrug. The formulation of such a
pharmaceutical composition can optionally enhance or achieve the
desired release profile.
[0173] In one embodiment, the invention provides methods for
treating a patient comprising administering a therapeutically
effective amount of an amphetamine prodrug or salt thereof, i.e.,
an amount sufficient to prevent, ameliorate, and/or eliminate the
symptoms of a disease. These methods can be used to treat any
disease that may benefit from amphetamine-type drugs including, but
not limited to: attention deficit disorders, e.g., ADD and ADHD,
and other learning disabilities; obesity; Alzheimer's disease,
amnesia, and other memory disorders and impairments; fibromyalgia;
fatigue and chronic fatigue; depression; epilepsy; obsessive
compulsive disorder (OCD); oppositional defiant disorder (ODD);
anxiety; resistant depression; stroke rehabilitation; Parkinson's
disease; mood disorder; schizophrenia; Huntington's disorder;
dementia, e.g., AIDS dementia and frontal lobe dementia; movement
disfunction; apathy; Pick's disease; Creutzfeldt-Jakob disease,
sleep disorders, e.g., narcolepsy, cataplexy, sleep paralysis,
cancer related fatigue, excessive daytime sleepiness in patients
suffering from obstructive sleep apnea and hypnagogic
hallucinations; conditions related to brain injury or neuronal
degeneration, e.g., multiple sclerosis, Tourette's syndrome, and
impotence; and nicotine dependence and withdrawal. Preferred
indications include ADD, ADHD, narcolepsy, and obesity, with ADHD
being most preferred.
[0174] In a further embodiment, the amphetamine prodrug to be
administered comprises amphetamine covalently bound to a peptide
comprising 1 to 10 amino acids. The prodrug can also be
administered in a pharmaceutical composition. A preferred prodrug
is L-lysine-d-amphetamine. A preferred prodrug salt is
L-lysine-d-amphetamine dimesylate.
[0175] Cancer Related Fatigue
[0176] In one embodiment, the prodrugs (or pharmaceutical salts
thereof) are administered to treat cancer related fatigue. The term
"cancer related fatigue" includes a patient's fatigue during the
period in which the patient is undergoing chemotherapy and/or
radiation treatment as well as a patient's fatigue while not
undergoing any such treatment (e.g., after a patient has undergone
chemotherapy and/or radiation treatment).
[0177] An effective amount of the prodrug (or pharmaceutically
acceptable salt thereof) is administered. For example, a daily dose
of 5-100 mg of the prodrug can be administered. According to one
embodiment, a daily dose of 5 or 10 mg of the prodrug is
administered and is optionally titrated every 1 to 7, e.g., 3-5
days (for example, 3 or 5 days), by 5 or 10 mg increments, to the
desired level of treatment. The maximum level of treatment is
preferably 70 mg. Alternatively, the dose can be held constant at 5
or 10 mg as well. Preferably, the daily dose is administered once
daily.
Excessive Daytime Sleepiness in Patients Suffering from Obstructive
Sleep Apnea
[0178] In one embodiment, the prodrugs (or pharmaceutical salts
thereof) are administered to treat excessive daytime sleepiness in
patients suffering from obstructive sleep apnea. An effective
amount of the prodrug (or pharmaceutically acceptable salt thereof)
is administered. For example, a daily dose of 5-100 mg of the
prodrug can be administered. According to one embodiment, a daily
dose of 5 or 10 mg of the prodrug is administered and is optionally
titrated every 3-5 days by 5 or 10 mg increments, to the desired
level of treatment. The maximum level of treatment is preferably 70
mg. Alternatively, the dose can be held constant at 5 or 10 mg as
well. Preferably, the daily dose is administered once daily.
[0179] Fatigue related to cancer, excessive daytime sleepiness in
patients suffering from obstructive sleep apnea, or fatigue related
to any other sleeping disorders can be measured by various
instruments. A non-exhaustive list of instruments to measure
fatigue include the Piper Fatigue Scale (Piper, B F, et al., Oncol.
Nurs. Forum, vol 25, pp. 677-84 (1998)), Lee Fatigue Scale (Lee, K
A, et al., Psychiatry Res., vol. 36, pp. 297-8 (1991)), Functional
Assessment of Cancer Therapy-Anemia/Fatigue (Yellen, S B, J. Pain
Symptom Manage., vol. 13, pp. 63-74 (1997)), Brief Fatigue
Inventory (Mendoza, T R, Cancer, vol 85, pp. 1186-96 (1999)),
Cancer Fatigue Scale (Okuyama, T, J. Pain Symptom Manage., vol. 19,
pp. 5-14 (2000)), Schwartz Cancer Fatigue Scale (Schwartz, A L,
Oncol. Nurs. Forum, vol 25, pp. 711-7 (1998)), Multidimensional
Fatigue Inventory (Smets, E M, J. Psychosom. Res., vol 39, pp.
315-25 (1995)), Fatigue Severity Scale (Krupp, L B, et al., Arch
Neurol., vol. 46, pp. 1121-3), Multidimensional Characterization of
Fatigue Measure (Vercoulen, J H, et al., Arch. Of Neurol., vol. 53,
pp. 642-49 (1996)), and The Pittsburgh Sleep Quality Index (PSQI)
(Buysse, D J, et al., Psychiatry Res., vol. 2, pp. 193-213
(1989)).
[0180] These studies employ self-reporting measures, which are
compared to controls lacking fatigue, result in a score which can
be compared before and after treatment to assess the improvement in
fatigue symptoms following treatment with the prodrug.
[0181] For example, the Pittsburgh Sleep Quality Index (PSQI) can
be used to determine whether the prodrug has an effect on cancer
related fatigue, or any other fatigue or sleep related disorder,
including excessive daytime sleepiness in patients suffering from
obstructive sleep apnea. The PSQI is a questionnaire which assesses
sleep quality and disturbances over a 1-month time interval.
Nineteen individual items generate seven component scores:
subjective sleep quality, sleep latency, sleep duration, habitual
sleep efficiency, sleep disturbances, use of sleeping medication,
and daytime dysfunction. The sum of the scores for these seven
components yields one global score (Buysse, D J, et al., Psychiatry
Res., vol. 2, pp. 193-213 (1989)).
[0182] Combination Therapies
[0183] The methods of treatment include combination therapies which
further comprise administering one or more therapeutic agents in
addition to administering an amphetamine prodrug (or
pharmaceutically acceptable salt thereof). In these embodiments,
the therapeutic agents can be administered serially or
together.
[0184] In one embodiment, the prodrugs of the present invention (or
pharmaceutically acceptable salts thereof) are used in combination
with another antidepressant for adjunctive antidepressant
therapy.
[0185] The active ingredients can be formulated into a single
dosage form, or they can be formulated together or separately among
multiple dosage forms. The active ingredients can be administered
simultaneously or sequentially in any order. Exemplary combination
therapies include the administration of the drugs listed in Table
1.
[0186] The prodrug (or pharmaceutical salts thereof) can be
administered in combination with an antidepressant to treat
depression or a depressive disorder, such as major depressive
disorder. The prodrug and antipressant can be can be administered
serially (in any order) or together (simultaneously). The active
ingredients can be formulated into a single dosage form, or they
can be formulated together or separately among multiple dosage
forms.
[0187] In the adjunctive antidepressant therapy embodiment, the
prodrugs of the present invention, and specifically the preferred
prodrug salt l-lysine-d-amphetamine dimesylate, can be paired with
various antidepressants for co-therapy, including, but not limited
to, serotonin norepinephrine reuptake inhibitors (SNRIs) (e.g.,
venlafaxine), selective serotonin reuptake inhibitors (SSRIs), and
tertiary amine tricyclic norepinephrine reuptake inhibitors. For
example, tertiary amine tricyclic norepinephrine reuptake
inhibitors such as amitriptyline, clomipramine, doxepin, imipramine
and (+)-trimipramine can be employed with l-lysine-d-amphetamine or
a pharmaceutically acceptable salt thereof (e.g., the dimesylate
salt).
[0188] In another adjunctive antidepressant therapy embodiment,
secondary amine tricyclics such as amoxapine, desipramine,
maprotiline, nortriptyline and protriptyline are paired with an
amphetamine prodrug for adjunctive antidepressant therapy. The
antidepressant can also be a selective serotonin reuptake
inhibitor, such as citalopram, escitalopram, fluoxetine,
fluvoxamine, paroxetine, or sertraline.
[0189] In yet another adjunctive antidepressant therapy embodiment,
a prodrug of the present invention is administered with an atypical
antidepressant. Non-limiting examples of such compounds include
atomoxetine, bupropion, duloxetine, nefazodone and trazadone.
[0190] Monoamine oxidase inhibitors can also be used in combination
with the prodrugs of the present invention for adjunctive
antidepressant therapy. Examples include phenelzine,
tranylcypromine and selegiline.
TABLE-US-00003 TABLE 1 Exemplary drug therapies contemplated for
use in combination with an amphetamine prodrug Condition Exemplary
drug class Specific exemplary drugs ADHD Amphetamine Ritalin .RTM.,
Dexedrine .RTM., Adderall .RTM., Cylert .RTM., Clonidine,
Guanfacine Alzheimer's disease Reminyl .RTM., Cognex .RTM., Aricept
.RTM., Exelon .RTM., Akatinol .RTM., Neotropin, Eldepryl .RTM.,
Estrogen, Clioquinol, Ibuprofen, Ginko Biloba Anxiety
Antidepressant (SSRI, Elavil, Asendin .RTM., benzodiazepine, MAOI),
Wellbutrin .RTM., Tegretol .RTM., anxiolytic Anafranil .RTM.,
Norpramine .RTM., Adapin .RTM., Sinequan .RTM., Tofranil .RTM.,
Epitol .RTM., Janimire .RTM., Pamelor .RTM., Ventyl .RTM., Aventyl
.RTM., Surmontil .RTM., Prozac .RTM., Luvox .RTM., Serzone .RTM.,
Paxil .RTM., Zoloft .RTM., Effexor .RTM., Xanax .RTM., Librium
.RTM., Klonopin .RTM., Valium .RTM., Zetran .RTM., Valrelease
.RTM., Dalmane .RTM., Ativan .RTM., Alzapam .RTM., Serax .RTM.,
Halcion .RTM., Aurorix .RTM., Manerix .RTM., Nardil .RTM., Parnate
.RTM.. Apathy Amisulpride, Olanzapine, Visperidone, Quetiapine,
Clozapine, Zotepine Cataplexy Xyrem .RTM. Dementia Thioridazine,
Haloperidol, Risperidone, Cognex .RTM., Aricept .RTM., Exelon .RTM.
Depression Antidepressant Fluoxetine (e.g., Prozac .RTM.), Zoloft
.RTM., Paxil .RTM., Reboxetine, Wellbutrin .RTM., Olanzapine,
Elavil .RTM., Totranil .RTM., Pamelor .RTM., Nardil .RTM., Parnate
.RTM., Desyrel .RTM., Effexor .RTM. Fatigue Benzodiazepine Anaprox
.RTM., Naprosen, Prozac .RTM., Zoloft .RTM., Paxil .RTM., Effexor
.RTM., Desyrel .RTM. Fibromyalgia Non-steroidal anti- Dilantin
.RTM., Carbatrol .RTM., inflammatory drugs Epitol .RTM., Tegretol
.RTM., Depacon .RTM., Depakote .RTM., Norpramin .RTM., Aventyl
.RTM., Pamelor .RTM., Elavil .RTM., Enovil .RTM., Adapin .RTM.,
Sinequan .RTM., Zonalon .RTM. Hallucinations Clozapine,
Risperidone, Zyprexa .RTM., Seroquel .RTM. Huntington's disorder
Haloperidol, Clonzepam Narcolepsy Modafinil (e.g., Provigil .RTM.),
Dexedrine .RTM., Ritalin .RTM. Mood disorder Thorazine .RTM.,
Haldol .RTM., Navane .RTM., Mellaril .RTM., Clozaril .RTM.,
Risperidone (e.g., Risperdal .RTM.), Olanzapine (e.g., Zyprexa
.RTM.), Clozapine Obsessive-compulsive SSRI Anafranil .RTM., Prozac
.RTM., disorder (OCD) Zoloft .RTM., Paxil .RTM., Luvox .RTM.
Oppositional defiant Clonidine, Risperidone, disorder (ODD) Zyprexa
.RTM., Wellbutrin .RTM., Parkinson's disease Levodopa, Parlodel
.RTM., Permax .RTM., Mirapex .RTM. Schizophrenia Clozapine, Zyprexa
.RTM., Seroquel .RTM., and Risperdal .RTM. Sleep paralysis
Clonazepam, Ritalin .RTM.
[0191] A "composition" refers broadly to any composition containing
one or more amphetamine prodrugs. The composition can comprise a
dry formulation, an aqueous solution, or a sterile composition.
Compositions comprising the compounds described herein may be
stored in freeze-dried form and may be associated with a
stabilizing agent such as a carbohydrate. In use, the composition
may be deployed in an aqueous solution containing salts, e.g.,
NaCl, detergents such as sodium dodecyl sulfate (SDS), and other
components.
[0192] In one embodiment, the amphetamine prodrug itself exhibits a
sustained release profile. Thus, the invention provides a
pharmaceutical composition exhibiting a sustained release profile
due to the amphetamine prodrug.
[0193] In another embodiment, a sustained release profile is
enhanced or achieved by including a hydrophilic polymer in the
pharmaceutical composition. Suitable hydrophilic polymers include,
but are not limited to, natural or partially or totally synthetic
hydrophilic gums such as acacia, gum tragacanth, locust bean gum,
guar gum, and karaya gum; cellulose derivatives such as methyl
cellulose, hydroxymethyl cellulose, hydroxypropylmethyl cellulose,
hydroxypropyl cellulose, hydroxyethyl cellulose, and carboxymethyl
cellulose; proteinaceous substances such as agar, pectin,
carrageen, and alginates; hydrophilic polymers such as
carboxypolymethylene; gelatin; casein; zein; bentonite; magnesium
aluminum silicate; polysaccharides; modified starch derivatives;
and other hydrophilic polymers known in the art. Preferably, the
hydrophilic polymer forms a gel that dissolves slowly in aqueous
acidic media thereby allowing the amphetamine prodrug to diffuse
from the gel in the stomach. Then when the gel reaches the higher
pH medium of the intestines, the hydrophilic polymer dissolves in
controlled quantities to allow further sustained release. Preferred
hydrophilic polymers are hydroxypropyl methylcelluloses such as
Methocel ethers, e.g., Methocel E10M.RTM. (Dow Chemical Company,
Midland, Mich.). One of ordinary skill in the art would recognize a
variety of structures, such as bead constructions and coatings,
useful for achieving particular release profiles. See, e.g., U.S.
Pat. No. 6,913,768.
[0194] In addition to the amphetamine prodrug, the pharmaceutical
compositions of the invention further comprise one or more
pharmaceutical additives. Pharmaceutical additives include a wide
range of materials including, but not limited to diluents and
bulking substances, binders and adhesives, lubricants, glidants,
plasticizers, disintegrants, carrier solvents, buffers, colorants,
flavorings, sweeteners, preservatives and stabilizers, and other
pharmaceutical additives known in the art. For example, in a
preferred embodiment, the pharmaceutical composition comprises
magnesium stearate. In another preferred embodiment, the
pharmaceutical composition comprises microcrystalline cellulose
(e.g., Avicel.RTM. PH-102), croscarmellose sodium, and magnesium
stearate. See, e.g., Table 62.
[0195] Diluents increase the bulk of a dosage form and may make the
dosage form easier to handle. Exemplary diluents include, but are
not limited to, lactose, dextrose, saccharose, cellulose, starch,
and calcium phosphate for solid dosage forms, e.g., tablets and
capsules; olive oil and ethyl oleate for soft capsules; water and
vegetable oil for liquid dosage forms, e.g., suspensions and
emulsions. Additional suitable diluents include, but are not
limited to, sucrose, dextrates, dextrin, maltodextrin,
microcrystalline cellulose (e.g., Avicel.RTM.), microfine
cellulose, powdered cellulose, pregelatinized starch (e.g., Starch
1500.RTM.), calcium phosphate dihydrate, soy polysaccharide (e.g.,
Emcosoy.RTM.), gelatin, silicon dioxide, calcium sulfate, calcium
carbonate, magnesium carbonate, magnesium oxide, sorbitol,
mannitol, kaolin, polymethacrylates (e.g., Eudragit.RTM.),
potassium chloride, sodium chloride, and talc. A preferred diluent
is microcrystalline cellulose (e.g., Avicel.RTM. PH-102). Preferred
ranges for the amount of diluent by weight percent include about
40% to about 90%, about 50% to about 85%, about 55% to about 80%,
about 50% to about 60%, and increments therein.
[0196] In embodiments where the pharmaceutical composition is
compacted into a solid dosage form, e.g., a tablet, a binder can
help the ingredients hold together. Binders include, but are not
limited to, sugars such as sucrose, lactose, and glucose; corn
syrup; soy polysaccharide, gelatin; povidone (e.g., Kollidon.RTM.,
Plasdone.RTM.); Pullulan; cellulose derivatives such as
microcrystalline cellulose, hydroxypropylmethyl cellulose (e.g.,
Methocel.RTM.), hydroxypropyl cellulose (e.g., Klucel.RTM.),
ethylcellulose, hydroxyethyl cellulose, carboxymethylcellulose
sodium, and methylcellulose; acrylic and methacrylic acid
co-polymers; carbomer (e.g., Carbopol.RTM.);
polyvinylpolypyrrolidine, polyethylene glycol (Carbowax.RTM.);
pharmaceutical glaze; alginates such as alginic acid and sodium
alginate; gums such as acacia, guar gum, and arabic gums;
tragacanth; dextrin and maltodextrin; milk derivatives such as
whey; starches such as pregelatinized starch and starch paste;
hydrogenated vegetable oil; and magnesium aluminum silicate.
[0197] For tablet dosage forms, the pharmaceutical composition is
subjected to pressure from a punch and dye. Among other purposes, a
lubricant can help prevent the composition from sticking to the
punch and dye surfaces. A lubricant can also be used in the coating
of a coated dosage form. Lubricants include, but are not limited
to, magnesium stearate, calcium stearate, zinc stearate, powdered
stearic acid, glyceryl monostearate, glyceryl palmitostearate,
glyceryl behenate, silica, magnesium silicate, colloidal silicon
dioxide, titanium dioxide, sodium benzoate, sodium lauryl sulfate,
sodium stearyl fumarate, hydrogenated vegetable oil, talc,
polyethylene glycol, and mineral oil. A preferred lubricant is
magnesium stearate. The amount of lubricant by weight percent is
preferably less than about 5%, more preferably 4%, 3%, 2%, 1.5%,
1%, or 0.5%, or increments therein.
[0198] Glidants can improve the flowability of non-compacted solid
dosage forms and can improve the accuracy of dosing. Glidants
include, but are not limited to, colloidal silicon dioxide, fumed
silicon dioxide, silica gel, talc, magnesium trisilicate, magnesium
or calcium stearate, powdered cellulose, starch, and tribasic
calcium phosphate.
[0199] Plasticizers include both hydrophobic and hydrophilic
plasticizers such as, but not limited to, diethyl phthalate, butyl
phthalate, diethyl sebacate, dibutyl sebacate, triethyl citrate,
acetyltriethyl citrate, acetyltributyl citrate, cronotic acid,
propylene glycol, castor oil, triacetin, polyethylene glycol,
propylene glycol, glycerin, and sorbitol. Plasticizers are
particularly useful for pharmaceutical compositions containing a
polymer and in soft capsules and film-coated tablets. In one
embodiment, the plasticizer facilitates the release of the
amphetamine prodrug from the dosage form.
[0200] Disintegrants can increase the dissolution rate of a
pharmaceutical composition. Disintegrants include, but are not
limited to, alginates such as alginic acid and sodium alginate,
carboxymethylcellulose calcium, carboxymethylcellulose sodium
(e.g., Ac-Di-Sol.RTM., Primellose.RTM.), colloidal silicon dioxide,
croscarmellose sodium, crospovidone (e.g., Kollidon.RTM.,
Polyplasdone.RTM.), polyvinylpolypyrrolidine (Plasone-XL.RTM.),
guar gum, magnesium aluminum silicate, methyl cellulose,
microcrystalline cellulose, polacrilin potassium, powdered
cellulose, starch, pregelatinized starch, sodium starch glycolate
(e.g., Explotab.RTM., Primogel.RTM.). Preferred disintegrants
include croscarmellose sodium and microcrystalline cellulose (e.g.,
Avicel.RTM. PH-102). Preferred ranges for the amount of
disintegrant by weight percent include about 1% to about 10%, about
1% to about 5%, about 2% to about 3%, and increments therein.
[0201] In embodiments where the pharmaceutical composition is
formulated for a liquid dosage form, the pharmaceutical composition
may include one or more solvents. Suitable solvents include, but
are not limited to, water; alcohols such as ethanol and isopropyl
alcohol; methylene chloride; vegetable oil; polyethylene glycol;
propylene glycol; and glycerin.
[0202] The pharmaceutical composition can comprise a buffer.
Buffers include, but are not limited to, lactic acid, citric acid,
acetic acid, sodium lactate, sodium citrate, and sodium
acetate.
[0203] Any pharmaceutically acceptable colorant can be used to
improve appearance or to help identify the pharmaceutical
composition. See 21 C.F.R., Part 74. Exemplary colorants include
D&C Red No. 28, D&C Yellow No. 10, FD&C Blue No. 1,
FD&C Red No. 40, FD&C Green #3, FD&C Yellow No. 6, and
edible inks. Preferred colors for gelatin capsules include white,
medium orange, and light blue.
[0204] Flavorings improve palatability and may be particularly
useful for chewable tablet or liquid dosage forms. Flavorings
include, but are not limited to maltol, vanillin, ethyl vanillin,
menthol, citric acid, fumaric acid, ethyl maltol, and tartaric
acid. Sweeteners include, but are not limited to, sorbitol,
saccharin, sodium saccharin, sucrose, aspartame, fructose,
mannitol, and invert sugar.
[0205] The pharmaceutical compositions of the invention can also
include one or more preservatives and/or stabilizers to improve
storagability. These include, but are not limited to, alcohol,
sodium benzoate, butylated hydroxy toluene, butylated
hydroxyanisole, and ethylenediamine tetraacetic acid.
[0206] Other pharmaceutical additives include gelling agents such
as colloidal clays; thickening agents such as gum tragacanth and
sodium alginate; wetting agents such as lecithin, polysorbates, and
laurylsulphates; humectants; antioxidants such as vitamin E,
caronene, and BHT; adsorbents; effervescing agents; emulsifying
agents, viscosity enhancing agents; surface active agents such as
sodium lauryl sulfate, dioctyl sodium sulfosuccinate,
triethanolamine, polyoxyethylene sorbitan, poloxalkol, and
quaternary ammonium salts; and other miscellaneous excipients such
as lactose, mannitol, glucose, fructose, xylose, galactose,
sucrose, maltose, xylitol, sorbitol, chloride, sulfate and
phosphate salts of potassium, sodium, and magnesium.
[0207] The pharmaceutical compositions can be manufactured
according to any method known to those of skill in the art of
pharmaceutical manufacture such as, for example, wet granulation,
dry granulation, encapsulation, direct compression, slugging, etc.
For instance, a pharmaceutical composition can be prepared by
mixing the amphetamine prodrug with one or more pharmaceutical
additives with an aliquot of liquid, preferably water, to form a
wet granulation. The wet granulation can be dried to obtain
granules. The resulting granulation can be milled, screened, and
blended with various pharmaceutical additives such as
water-insoluble polymers and additional hydrophilic polymers. In
one embodiment, an amphetamine prodrug is mixed with a hydrophilic
polymer and an aliquot of water, then dried to obtain granules of
amphetamine prodrug encapsulated by hydrophilic polymer.
[0208] After granulation, the pharmaceutical composition is
preferably encapsulated, e.g., in a gelatin capsule. The gelatin
capsule can contain, for example, kosher gelatin, titanium dioxide,
and optional colorants. Alternatively, the pharmaceutical
composition can be tableted, e.g., compressed and optionally coated
with a protective coating that dissolves or disperses in gastric
juices.
[0209] The pharmaceutical compositions of the invention can be
administered by a variety of dosage forms. Any
biologically-acceptable dosage form known in the art, and
combinations thereof, are contemplated. Examples of preferred
dosage forms include, without limitation, tablets including
chewable tablets, film-coated tablets, quick dissolve tablets,
effervescent tablets, multi-layer tablets, and bi-layer tablets;
caplets; powders including reconstitutable powders; granules;
dispersible granules; particles; microparticles; capsules including
soft and hard gelatin capsules; lozenges; chewable lozenges;
cachets; beads; liquids; solutions; suspensions; emulsions;
elixirs; and syrups.
[0210] The pharmaceutical composition is preferably administered
orally. Oral administration permits the maximum release of
amphetamine, provides sustained release of amphetamine, and
maintains abuse resistance. Preferably, the amphetamine prodrug
releases the amphetamine over a more extended period of time as
compared to administering unbound amphetamine.
[0211] Oral dosage forms can be presented as discrete units, such
as capsules, caplets, or tablets. In a preferred embodiment, the
invention provides a solid oral dosage form comprising an
amphetamine prodrug that is smaller in size compared to a solid
oral dosage form containing a therapeutically equivalent amount of
unbound amphetamine. In one embodiment, the oral dosage form
comprises a gelatin capsule of size 2, size 3, or smaller (e.g.,
size 4). The smaller size of the amphetamine prodrug dosage forms
promotes ease of swallowing.
[0212] Soft gel or soft gelatin capsules may be prepared, for
example, by dispersing the formulation in an appropriate vehicle
(e.g., vegetable oil) to form a high viscosity mixture. This
mixture then is encapsulated with a gelatin based film. The
industrial units so formed are then dried to a constant weight.
[0213] Chewable tablets can be prepared by mixing the amphetamine
prodrug with excipients designed to form a relatively soft,
flavored tablet dosage form that is intended to be chewed.
Conventional tablet machinery and procedures (e.g., direct
compression, granulation, and slugging) can be utilized.
[0214] Film-coated tablets can be prepared by coating tablets using
techniques such as rotating pan coating methods and air suspension
methods to deposit a contiguous film layer on a tablet.
[0215] Compressed tablets can be prepared by mixing the amphetamine
prodrug with excipients that add binding qualities. The mixture can
be directly compressed, or it can be granulated and then
compressed.
[0216] The pharmaceutical compositions of the invention can
alternatively be formulated into a liquid dosage form, such as a
solution or suspension in an aqueous or non-aqueous liquid. The
liquid dosage form can be an emulsion, such as an oil-in-water
liquid emulsion or a water-in-oil liquid emulsion. The oils can be
administered by adding the purified and sterilized liquids to a
prepared enteral formula, which then is placed in the feeding tube
of a patient who is unable to swallow.
[0217] For oral administration, fine powders or granules containing
diluting, dispersing, and/or surface-active agents can be presented
in a draught, in water or a syrup, in capsules or sachets in the
dry state, in a non-aqueous suspension wherein suspending agents
may be included, or in a suspension in water or a syrup. Liquid
dispersions for oral administration can be syrups, emulsions, or
suspensions. The syrups, emulsions, or suspensions can contain a
carrier, for example, a natural gum, agar, sodium alginate, pectin,
methylcellulose, carboxymethylcellulose, saccharose, saccharose
with glycerol, mannitol, sorbitol, and polyvinyl alcohol.
[0218] The dose range of the amphetamine prodrug for humans will
depend on a number of factors including the age, weight, and
condition of the patient. Tablets and other dosage forms provided
in discrete units can contain a daily dose, or an appropriate
fraction thereof, of one or more amphetamine prodrugs. The dosage
form can contain a dose of about 2.5 mg to about 500 mg, about 10
mg to about 250 mg, about 10 mg to about 100 mg, about 25 mg to
about 75 mg, or increments therein of one or more of the
amphetamine prodrugs. In a preferred embodiment, the dosage form
contains 30 mg, 50 mg, or 70 mg of an amphetamine prodrug.
[0219] The dosage form can utilize any one or any combination of
known release profiles including, but not limited to immediate
release, extended release, pulse release, variable release,
controlled release, timed release, sustained release, delayed
release, and long acting.
[0220] The pharmaceutical compositions of the invention can be
administered in a partial, i.e., fractional dose, one or more times
during a 24 hour period. Fractional, single, double, or other
multiple doses can be taken simultaneously or at different times
during a 24 hour period. The doses can be uneven doses with regard
to one another or with regard to the individual components at
different administration times. Preferably, a single dose is
administered once daily. The dose can be administered in a fed or
fasted state.
[0221] The dosage units of the pharmaceutical composition can be
packaged according to market need, for example, as unit doses,
rolls, bulk bottles, blister packs, and so forth. The
pharmaceutical package, e.g., blister pack, can further include or
be accompanied by indicia allowing individuals to identify the
identity of the pharmaceutical composition, the prescribed
indication (e.g., ADHD), and/or the time periods (e.g., time of
day, day of the week, etc.) for administration. The blister pack or
other pharmaceutical package can also include a second
pharmaceutical product for combination therapy.
[0222] It will be appreciated that the pharmacological activity of
the compositions of the invention can be demonstrated using
standard pharmacological models that are known in the art.
Furthermore, it will be appreciated that the inventive compositions
can be incorporated or encapsulated in a suitable polymer matrix or
membrane for site-specific delivery, or can be functionalized with
specific targeting agents capable of effecting site specific
delivery. These techniques, as well as other drug delivery
techniques, are well known in the art.
[0223] Any feature of the above-describe embodiments can be used in
combination with any other feature of the above-described
embodiments.
[0224] In order to facilitate a more complete understanding of the
invention, Examples are provided below. However, the scope of the
invention is not limited to specific embodiments disclosed in these
Examples, which are for purposes of illustration only.
EXAMPLES
[0225] The following abbreviations are used in the Examples and
throughout the specification: [0226]
Lys-Amp=L-lysine-d-amphetamine, Lysine-Amphetamine, K-Amp,
K-amphetamine, or 2,6-diaminohexanoic
acid-(1-methyl-2-phenylethyl)-amide, lisdexamphetamine or
Lisdexamfetamine [0227] Phe-Amp=Phenylalanine-Amphetamine, F-Amp,
or 2-amino-3-phenylpropanoic acid-(1-methyl-2-phenylethyl)-amide
[0228] Ser-Amp=Serine-Amphetamine, S-Amp, or
2-amino-3-hydroxylpropanoic acid-(1-methyl-2-phenylethyl)-amide,
[0229] Gly.sub.3-Amp=GGG-Amphetamine, GGG-Amp, or
2-amino-N-({[(1-methyl-2-phenyl-ethylcarbornyl)-methyl]-carbornyl}-methyl-
)-acetamide [0230] BOC=t-butyloxycarbonyl [0231]
CMC=carboxymethylcellulose [0232] DIPEA=di-isopropyl ethyl amine
[0233] mp=melting point [0234] NMR=nuclear magnetic resonance
[0235] OSu=hydroxysuccinimido ester
[0236] Throughout the Examples, unless otherwise specified, doses
are described as the amount of d-amphetamine base. Exemplary
conversions are provided in Table 2.
TABLE-US-00004 TABLE 2 Conversion of d-amphetamine doses (mg)
L-lysine-d-amphetamine dimesylate d-amphetamine d-amphetamine
sulfate (29.5% d-amphetamine) base (72.8% d-amphetamine) 5.08 1.5
2.06 10.17 3 4.12 20.34 6 8.24 40.68 12 16.48 101.69 30 41.21
203.39 60 82.42 25.00 7.375 10.13 75.00 22.125 30.39 70.00 20.65
28.37 50.00 14.75 20.26 30.00 8.85 12.16
[0237] X-Ray Powder Diffraction (XRPD) ANALYSES
[0238] XRPD patterns were collected using a PANalytical X'Pert Pro
diffractometer. The specimens were analyzed using Cu radiation
produced using an Optix long fine-focus source. An alleptically
graded multilayer mirror was used to focus the Cu K .alpha. X-rays
of the source through the specimen and onto the detector. Each
specimen was sandwiched between 3 .mu.m thick films, analyzed in
transmission geometry, and rotated to optimize orientation
statistics. A beam-stop and helium purge was used to minimize the
background generated by air scattering. Soller slits were used for
the incident and diffracted beams to minimize axial divergence.
Diffraction patterns were collected using a scanning
position-sensitive detector (X-celerator) located 240 mm from the
specimen. The data-acquisition parameters of each diffraction
pattern are given in the respective examples, below. Prior to the
analysis a silicon specimen (NIST standard reference material 640c)
was analyzed to verify the position of the silicon 111 peak.
[0239] The XRPD pattern of lisdexamphetamine dimesylate was indexed
using proprietary software--PatternMatch 2.4.0, provided by SSCI
(West Lafayette, Ind.). The solution was further refined via Pawley
refinement using DASH version 3.0, provided by Cambridge
Crystallographic Data Centre (Cambridge, UK). The indexed solution
was verified and illustrated using CheckCell version Nov. 1, 2004
(ccp14.ac.uk/tutorial/lmgp/index.html).
[0240] DSC Analyses
[0241] DSC was performed using a TA Instruments differential
scanning calorimeter 2920 and Q2000. The respective sample was
placed into an aluminum DSC pan, and the weight accurately
recorded. The pan was covered with a lid and lift uncrimped. The
sample cell was equilibrated at 25.degree. C. and heated under a
nitrogen purge at a rate of 10.degree. C./min, up to a final
temperature of 250.degree. C. Indium metal was used as the
calibration standard. Reported temperatures are at the transition
maxima and peak onsets.
[0242] FT-Raman Spectroscopy
[0243] FT-Raman spectra were acquired on a FT-Raman 960
spectrometer (Thermo Nicolet, Woburn, Mass.). This spectrometer
uses an excitation wavelength of 1064 nm. Approximately 0.5 W of
Nd:YVO.sub.4 laser power was used to irradiate the respective
sample. The Raman spectra were measured with a germanium (Ge)
detector. The samples were prepared for analysis by placing the
material in a glass tube and positioning the tube in a gold-coated
tube holder in the accessory. A total of 256 sample scans were
collected from 3600-100 cm.sup.-1 at a spectral resolution of 4
cm.sup.-1, using Happ-Genzel apodization. Wavelength calibration
was performed using sulfur and cyclohexane.
[0244] Particle Size Determination
[0245] The micrometer bar size was established for both the SEM and
light microscope(s) in standard resolution by comparison with a
NIST traceable calibration standard. The software for each
instrument then determines the size of each pixel in each image,
and uses this information for comparison with objects shown in the
photomicrograph. The comparison takes into account the
magnification of the image and calculates the true size of the
object, not the magnified size.
Example 1
General Synthesis of Peptide Amphetamine Conjugates
[0246] Peptide conjugates were synthesized by the general method
described in FIG. 1. An iterative approach can be used to identify
favorable conjugates by synthesizing and testing single amino acid
conjugates, and then extending the peptide one amino acid at a time
to yield dipeptide and tripeptide conjugates, etc. The parent
single amino acid prodrug candidate may exhibit more or less
desirable characteristics than its di- or tripeptide offspring
candidates. The iterative approach can quickly suggest whether
peptide length influences bioavailability.
General Synthesis of Single Amino Acid Amphetamine Conjugates
[0247] To a solution of a protected amino acid succinimidyl ester
(2.0 eq) in 1,4-dioxane (30 mL) was added d-amphetamine sulfate
(1.0 eq) and NMM (4.0 eq). The resulting mixture was allowed to
stir for 20 h at 20.degree. C. Water (10 mL) was added, and the
solution was stirred for 10 minutes prior to removing solvents
under reduced pressure. The crude product was dissolved in EtOAc
(100 mL) and washed with 2% AcOH.sub.aq (3.times.100 mL), saturated
NaHCO.sub.3 solution (2.times.50 mL), and brine (1.times.100 mL).
The organic extract was dried over MgSO.sub.4, filtered, and
evaporated to dryness to afford the protected amino acid
amphetamine conjugate. This intermediate was directly deprotected
by adding 4 N HCl in 1,4-dioxane (20 mL). The solution was stirred
for 20 h at 25.degree. C. The solvent was evaporated, and the
product dried in vacuum to afford the corresponding amino acid
amphetamine hydrochloride conjugate. The syntheses of exemplary
single amino acid conjugates are depicted in FIG. 2-FIG. 6.
General Synthesis of Dipeptide Amphetamine Conjugates
[0248] To a solution of a protected dipeptide succinimidyl ester
(1.0 eq) in 1,4-dioxane was added amphetamine sulfate (2.0 eq) and
NMM (4.0 eq). The resulting mixture was stirred for 20 h at
25.degree. C. Solvents were removed under reduced pressure.
Saturated NaHCO.sub.3 solution (20 mL) was added, and the
suspension was stirred for 30 min. IPAC (100 mL) was added, and the
organic layer was washed with 2% AcOH.sub.aq (3.times.100 mL) and
brine (2.times.100 mL). The organic extract was dried over
Na.sub.2SO.sub.4, and the solvent was evaporated to dryness to
yield the protected dipeptide amphetamine conjugate. The protected
dipeptide conjugate was directly deprotected by adding 4 N HCl in
1,4-dioxane (20 mL), and the solution stirred for 20 h at
25.degree. C. The solvent was evaporated, and the product was dried
in vacuum to afford the corresponding dipeptide amphetamine
hydrochloride conjugate.
General Synthesis of Tripeptide Amphetamine Conjugates
[0249] An amino acid conjugate was synthesized and deprotected
according to the general procedure described above. To a solution
of the amino acid amphetamine hydrochloride (1.0 eq) in dioxane (20
mL) was added NMM (5.0 eq) and a protected dipeptide succinate
(1.05 eq). The solution was stirred for 20 h at 25.degree. C. The
solvent was removed under reduced pressure. Saturated NaHCO.sub.3
solution (20 mL) was added, and the suspension was stirred for 30
min. IPAC (100 mL) was added, and the organic layer was washed with
2% AcOH.sub.aq (3.times.100 mL) and brine (2.times.100 mL). The
organic extract was dried over Na.sub.2SO.sub.4, and the solvent
was evaporated to dryness to yield the protected tripeptide
amphetamine. Deprotection was directly carried out by adding 4 N
HCl in 1,4-dioxane (20 mL). The mixture was stirred for 20 h at
25.degree. C., the solvent was evaporated, and the product was
dried in vacuum to afford the respective tripeptide amphetamine
hydrochloride conjugate.
[0250] The hydrochloride conjugates required no further
purification, but many of the deprotected hydrochloride salts were
hygroscopic and required special handling during analysis and
subsequent in vivo testing.
Example 2
Synthesis of L-lysine-d-amphetamine
[0251] L-lysine-d-amphetamine was synthesized by the following
methods.
[0252] a. Preparation of HCl Salt (see FIG. 3)
[0253] i. Coupling
TABLE-US-00005 Molar Reagents MW Weight mmoles Equivalents
d-amphetamine free base 135.2 4.75 g 35.13 1 Boc-Lys(Boc)-OSu 443.5
15.58 g 35.13 1 Di-iPr-Et-Amine 129 906 mg 7.03 0.2, d = 0.74, 1.22
mL 1,4-Dioxane -- 100 mL -- --
[0254] To a solution of Boc-Lys(Boc)-OSu (15.58 g, 35.13 mmol) in
dioxane (100 mL) under an inert atmosphere was added d-amphetamine
free base (4.75 g, 35.13 mmol) and DIPEA (0.9 g, 1.22 mL, 7.03
mmol). The resulting mixture was allowed to stir at room
temperature overnight. Solvent and excess base were then removed
using reduced pressure evaporation. The crude product was dissolved
in ethyl acetate and loaded on to a flash column (7 cm wide, filled
to 24 cm with silica) and eluted with ethyl acetate. The product
was isolated, the solvent reduced by rotary evaporation, and the
purified protected amide was dried by high-vac to obtain a white
solid. .sup.1H NMR (DMSO-d.sub.6) .delta. 1.02-1.11 (m, 2H, Lys
.gamma.-CH.sub.2), .delta. 1.04 (d, 3H, Amp .alpha.-CH.sub.3),
.delta. 1.22-1.43 (m, 4H, Lys-.beta. and .delta.-CH.sub.2), .delta.
1.37 (18H, Boc, 6.times.CH.sub.3), .delta. 2.60-2.72 (2H, Amp
CH.sub.2), .delta. 3.75-3.83, (m, 1H, Lys .alpha.-H) .delta.
3.9-4.1 (m, 1H, Amp .alpha.-H), .delta. 6.54-6.61 (d, 1H, amide
NH), .delta. 6.7-6.77 (m, 1H, amide NH), .delta. 7.12-7.29 (m, 5H,
ArH), .delta. 7.65-7.71 (m, 1, amide NH); mp=86-88.degree. C.
[0255] ii. Deprotection
TABLE-US-00006 Molar Reagents MW Weight mmoles Equivalents 4M HCl
in dioxane 4 mmol/mL 50 mL 200 6.25 Boc-Lys(Boc)-Amp 463.6 14.84 g
32 1 1,4-Dioxane -- 50 mL -- --
[0256] The protected amide was dissolved in 50 mL of anhydrous
dioxane and stirred while 50 mL (200 mmol) of 4M HCl/dioxane was
added and stirred at room temperature overnight. The solvents were
then reduced by rotary evaporation to afford a viscous oil.
Addition of 100 mL MeOH followed by rotary evaporation resulted in
a golden colored solid material that was further dried by storage
at room temperature under high vacuum. .sup.1H NMR (DMSO-d.sub.6)
.delta. 0.86-1.16 (m, 2H, Lys .gamma.-CH.sub.2), .delta. 1.1 (d,
3H, Amp .alpha.-CH.sub.3), .delta. 1.40-1.56 (m, 4H, Lys-.beta. and
.delta.-CH.sub.2), .delta. 2.54-2.78 (m, 2H, Amp CH.sub.2, 2H, Lys
.epsilon.-CH.sub.2), 3.63-3.74 (m, 1H, Lys .alpha.-H), .delta.
4.00-4.08 (m, 1H, Amp .alpha.-H), .delta. 7.12-7.31 (m, 5H, Amp
ArH), .delta. 8.13-8.33 (d, 3H, Lys amine) .delta. 8.70-8.78 (d,
1H, amide NH); mp=120-122.degree. C.
[0257] b. Preparation of Mesylate Salt (and See FIG. 2)
[0258] Similarly, the mesylate salt of the peptide conjugate can be
prepared by using methanesulfonic acid in the deprotection step as
described in further detail below.
[0259] i. Coupling
[0260] A 72-L round-bottom reactor was equipped with a mechanical
stirrer, digital thermocouple, and addition funnel and purged with
nitrogen. The vessel was charged with Boc-Lys(Boc)-OSu (3.8 kg,
8.568 mol, 1.0 eq) and 1,4-dioxane (20.4 L), and the resulting
turbid solution was stirred at 20.+-.5.degree. C. for 10 min. To
the mixture was added N-methylmorpholine (950 g, 9.39 mol, 1.09 eq)
over a period of 1 min, and the mixture was stirred for 10 min. To
the slightly turbid reaction mixture was then added a solution of
dextro-amphetamine (1.753 kg, 12.96 mol, 1.51 eq) in 1,4-dioxane
(2.9 L) over a period of 30 min, while cooling the reactor
externally with an ice/water bath. The internal temperature was
kept below 25.degree. C. during the addition. At the end of the
addition, a thick white precipitate appeared. The addition funnel
was rinsed with 1,4-dioxane (2.9 L) into the reactor, and the
reaction mixture was stirred at 22.+-.3.degree. C. TLC monitoring
30 min. after completed addition showed no more remaining
Boc-Lys(Boc)-Osu, and the reaction was quenched with DI H.sub.2O
(10 L). The mixture was stirred for 1 h at ambient temperature and
then concentrated under reduced pressure to afford a dense, white
solid.
[0261] For the extractions, two solutions were prepared: an acetic
acid/salt solution: NaCl (15 kg) and glacial acetic acid (2 kg) in
DI H.sub.2O (61 L), and a bicarbonate solution: NaHCO.sub.3 (1.5
kg) in DI H.sub.2O (30 L).
[0262] The solid was re-dissolved in IPAC (38 L) and acetic
acid/salt solution (39 kg) and transferred into a 150-L reactor.
The layers were mixed for 10 min. and then allowed to separate. The
organic layer was drained and washed with another portion (39 kg)
of acetic acid/salt solution, followed by a wash with bicarbonate
solution (31.5 kg). All phase separations occurred within 5 min. To
the organic solution was then added silica-gel (3.8 kg; Silica-gel
60). The resulting slurry was stirred for 45 min. and then filtered
through filter paper. The filter-cake was washed with IPAC
(5.times.7.6 L). The filtrate and washes were analyzed by TLC, and
it was determined that all contained product. The filtrate and
washes were combined and concentrated under reduced pressure to
afford the crude product as a white solid.
[0263] ii. Deprotection
[0264] A 45-L carboy was charged with di-Boc-Lys-Amp (3.63 kg,
7.829 mol) and 1,4-dioxane (30.8 L, 8.5 vol), and the mixture was
stirred rapidly under nitrogen for 30 min. The resulting solution
was filtered, and the filter-cake was rinsed with 1,4-dioxane
(2.times.1.8 L).
[0265] The filtrates were then transferred into a 72-L round-bottom
flask, which was equipped with a mechanical stirrer, digital
thermocouple, nitrogen inlet and outlet, and 5 L addition funnel.
The temperature of the reaction mixture was regulated at
21.+-.3.degree. C. with a water bath. To the clear, slightly yellow
solution was added methanesulfonic acid (3.762 kg, 39.15 mol, 5 eq)
over a period of 1 h while keeping the internal temperature at
21.+-.3.degree. C. Approximately 1 h after completed addition, a
white precipitate started to appear. The mixture was stirred at
ambient temperature for 20.5 h, after which HPLC monitoring showed
the disappearance of all starting material. The mixture was
filtered through filter-paper, and the reaction vessel was rinsed
with 1,4-dioxane (3.6 L, 1 vol). The filter-cake was washed with
dioxane (3.times.3.6 L) and dried with a rubber dam for 1 h. The
material was then transferred to drying trays and dried in a vacuum
oven at 55.degree. C. for .about.90 h. This afforded Lys-Amp
dimesylate [3.275 kg, 91.8% yield; >99% (AUC)] as a white
solid.
Example 3
Synthesis of Ser-Amp
[0266] Ser-Amp was synthesized by a similar method (see FIG. 4)
except the amino acid starting material was Boc-Ser(O-tBu)-OSu and
the deprotection was done using a solution of trifluoroacetic acid
instead of HCl.
Example 4
Synthesis of Phe-Amp
[0267] Phe-Amp was synthesized by a similar method (see FIG. 5)
except the amino acid starting material was Boc-Phe-OSu.
[0268] Phe-Amp hydrochloride: hygroscopic; .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 8.82 (d, J=8.0 Hz, 1H), 8.34 (bs, 3H),
7.29-7.11 (m, 10H), 3.99 (m, 2H), 2.99 (dd, J=13.6, 6.2 Hz, 1H),
2.88 (dd, J=13.6, 7.2 Hz, 1H), 2.64 (dd, J=13.2, 7.6 Hz, 1H), 2.53
(m, 1H), 1.07 (d, J=6.4 Hz, 3H); .sup.13C NMR (100 MHz,
DMSO-d.sub.6): .delta. 167.31, 139.27, 135.49, 130.05, 129.66,
128.78, 128.61, 127.40, 126.60, 53.83, 47.04, 42.15, 37.27, 20.54;
HRMS: (ESI) for C.sub.18H.sub.23N.sub.2O (M+H).sup.+: calcd,
283.1810: found, 283.1806.
Example 5
Synthesis of Gly.sub.3-Amp
[0269] Gly.sub.3-Amp was synthesized by a similar method (see FIG.
6) except the amino acid starting material was Boc-GGG-OSu.
[0270] Gly.sub.3-Amp hydrochloride: mp 212-214.degree. C.; .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 7.28 (m, 5H), 3.96 (m, 1H),
3.86 (m, 2H), 3.66 (m, 4H), 2.76 (m, 1H), 2.75 (m, 1H), 1.02 (d,
J=6.8 Hz, 3H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta. 168.91,
168.14, 166.85, 139.45, 129.60, 128.60, 126.48, 46.60, 42.27,
20.30. HRMS: (ESI) for C.sub.15H.sub.22N.sub.4O.sub.3Na
(M+Na).sup.+: calcd, 329.1590: found, 329.1590.
Example 6
Pharmacokinetics of L-lysine-d-amphetamine diHCl Compared to
d-amphetamine Sulfate (ELISA Analysis)
[0271] Male Sprague-Dawley rats were provided water ad libitum,
fasted overnight, and dosed by oral gavage L-lysine-d-amphetamine
diHCl or d-amphetamine sulfate. In all studies, doses contained
equivalent amounts of d-amphetamine base. Plasma d-amphetamine
concentrations were measured by ELISA (Amphetamine Ultra, 109319,
Neogen, Corporation, Lexington, Ky.). The assay is specific for
d-amphetamine with only minimal reactivity (0.6%) of the major
d-amphetamine metabolite (para-hydroxy-d-amphetamine) occurring.
L-lysine-d-amphetamine diHCl was also determined to be essentially
unreactive in the ELISA (<1%).
[0272] Mean (n=4) plasma concentration curves of d-amphetamine or
L-lysine-d-amphetamine diHCl are shown in FIG. 7. Extended release
was observed in all four L-lysine-d-amphetamine diHCl dosed
animals, and C.sub.max was substantially decreased as compared to
animals dosed with d-amphetamine sulfate. Plasma d-amphetamine
concentrations of individual animals for d-amphetamine or
L-lysine-d-amphetamine diHCl are shown in Table 3. The mean plasma
d-amphetamine concentrations are shown in Table 4. The time to peak
concentration for L-lysine-d-amphetamine diHCl was similar to that
of d-amphetamine. Pharmacokinetic parameters for oral
administration of d-amphetamine or L-lysine-d-amphetamine diHCl are
summarized in Table 5.
TABLE-US-00007 TABLE 3 Plasma concentrations of d-amphetamine from
individual animals orally administered d-amphetamine or
L-lysine-d-amphetamine diHCl (3 mg/kg d- amphetamine base) Time
d-amphetamine (ng/ml) L-lysine-d-amphetamine (ng/ml) (hours) Rat #1
Rat #2 Rat #3 Rat #4 Rat #1 Rat #2 Rat #3 Rat #4 0.5 144 157 101
115 52 62 74 44 1 152 78 115 78 48 72 79 57 1.5 85 97 117 95 42 62
76 53 3 34 45 72 38 61 60 71 43 5 20 14 12 15 49 33 44 22 8 3 3 2 2
15 14 12 8
TABLE-US-00008 TABLE 4 Mean plasma concentrations of d-amphetamine
following oral administration of d-amphetamine or
L-lysine-d-amphetamine Plasma d-amphetamine Concentrations (ng/ml)
d-amphetamine L-lysine-d-amphetamine Hours Mean .+-.SD CV Mean
.+-.SD CV 0.5 129 25 20 58 13 22 1 106 35 33 64 14 22 1.5 99 13 14
58 14 25 3 47 17 36 59 11 19 5 15 4 24 37 12 32 8 2 1 35 12 3
24
TABLE-US-00009 TABLE 5 Pharmacokinetic parameters of d-amphetamine
following oral administration of d-amphetamine or
L-lysine-d-amphetamine Mean AUC (0-8 h) Percent C.sub.max Percent
Peak Percent Drug ng h/mL Amphetamine (ng/ml) Amphetamine (ng/ml)
Amphetamine Amphetamine 341 .+-. 35 100 111 .+-. 27 100 129 100
Lys-Amp 333 .+-. 66 98 61 .+-. 13 55 64 50
[0273] This example illustrates that when lysine is conjugated to
the active agent amphetamine, the peak levels of amphetamine are
decreased while bioavailability is maintained approximately equal
to amphetamine. The bioavailability of amphetamine released from
L-lysine-d-amphetamine is similar to that of amphetamine sulfate at
the equivalent dose; thus L-lysine-d-amphetamine maintains its
therapeutic value. The gradual release of amphetamine from
L-lysine-d-amphetamine and decrease in peak levels reduce the
possibility of overdose.
Example 7
Oral Bioavailability of L-lysine-d-amphetamine Dimesylate at
Various Doses
[0274] a. Doses Approximating Therapeutic Human Doses (1.5, 3, and
6 mg/kg)
[0275] Mean (n=4) plasma concentration curves of d-amphetamine vs.
L-lysine-d-amphetamine are shown for rats orally administered 1.5,
3, and 6 mg/kg in FIG. 8, FIG. 9, and FIG. 10, respectively.
Extended release was observed at all three therapeutic doses for
L-lysine-d-amphetamine dosed animals. The mean plasma
concentrations for 1.5, 3, and 6 mg/kg are shown in Table 6, Table
7, and Table 8, respectively. Pharmacokinetic parameters for oral
administration of d-amphetamine vs. L-lysine-d-amphetamine at the
various doses are summarized in Table 9.
TABLE-US-00010 TABLE 6 Mean plasma concentrations of d-amphetamine
vs. L-lysine-d- amphetamine following oral administration (1.5
mg/kg) Plasma Amphetamine Concentrations (ng/ml) d-amphetamine
L-lysine-d-amphetamine Hours Mean .+-.SD CV Mean .+-.SD CV 0 0 0 0
0 0 0 0.25 103 22 21 31 11 37 0.5 126 20 16 51 23 45 1 101 27 27 68
23 34 1.5 116 28 24 72 10 14 3 66 13 20 91 5 5 5 40 7 18 75 16 22 8
17 2 15 39 13 34
TABLE-US-00011 TABLE 7 Mean plasma concentrations of d-amphetamine
vs. L-lysine-d- amphetamine following oral administration (3 mg/kg)
Plasma Amphetamine Concentrations (ng/ml) d-amphetamine
L-lysine-d-amphetamine Hours Mean .+-.SD CV Mean .+-.SD CV 0 0 0
0.25 96 41 43 51 49 97 0.5 107 49 46 36 35 96 1 121 17 14 81 44 54
1.5 120 33 27 97 32 33 3 91 30 33 88 13 15 5 62 22 36 91 21 23 8 19
6 33 46 16 34
TABLE-US-00012 TABLE 8 Mean plasma concentrations of d-amphetamine
vs. L-lysine-d- amphetamine following oral administration (6 mg/kg)
Plasma Amphetamine Concentrations (ng/ml) d-amphetamine
L-lysine-d-amphetamine Hours Mean .+-.SD CV Mean .+-.SD CV 0 0 0
0.25 204 14 7 74 38 51 0.5 186 9 5 106 39 37 1 167 12 7 133 33 24
1.5 161 24 15 152 22 15 3 111 29 26 157 15 10 5 78 9 11 134 18 13 8
35 5 15 79 12 15
TABLE-US-00013 TABLE 9 Pharmacokinetic parameters of d-amphetamine
following oral administration of d-amphetamine or
L-lysine-d-amphetamine 1.5 mg/kg 3 mg/kg 6 mg/kg Parameter Amp
K-Amp Amp K-Amp Amp K-Amp AUC 481 538 587 614 807 1005 (ng h/ml)
Percent 100 112 100 105 100 125 C.sub.max (ng/ml) 133 93 141 104
205 162 Percent 100 70 100 74 100 79 T.sub.max (hours) 0.938 3.5 1
1.56 0.563 2.625 Percent 100 373 100 156 100 466
[0276] b. Increased Doses (12, 30, and 60 mg/kg)
[0277] Mean (n=4) plasma concentration curves of d-amphetamine vs.
L-lysine-d-amphetamine are shown for rats orally administered 12,
30, and 60 mg/kg. At these higher doses, the bioavailability of
L-lysine-d-amphetamine was markedly decreased as compared to
d-amphetamine.
TABLE-US-00014 TABLE 10 Mean plasma concentrations of d-amphetamine
vs. L-lysine-d- amphetamine following oral administration (12
mg/kg) Plasma Amphetamine Concentrations (ng/ml) d-amphetamine
L-lysine-d-amphetamine Hours Mean .+-.SD CV Mean .+-.SD CV 0 NA NA
NA NA NA NA 0.25 530 279 53 53 34 64 0.5 621 76 12 99 32 33 1 512
91 18 220 77 35 1.5 519 113 22 224 124 55 3 376 149 40 300 153 51 5
314 123 39 293 153 52 8 103 64 63 211 45 22
TABLE-US-00015 TABLE 11 Mean plasma concentrations of d-amphetamine
vs. L-lysine-d- amphetamine following oral administration (30
mg/kg) Plasma Amphetamine Concentrations (ng/ml) L-lysine-d-
d-amphetamine amphetamine Hours Mean .+-.SD CV Mean .+-.SD CV 0 NA
NA NA NA NA NA 0.25 2,036 1,262 62 29 16 54 0.5 2,583 1,465 57 88
29 34 1 3,162 772 24 328 30 9 1.5 3,445 191 6 368 99 27 3 2,620 72
3 620 79 13 5 1,535 21 1 730 169 23 8 164 52 32 NA NA NA
TABLE-US-00016 TABLE 12 Mean plasma concentrations of d-amphetamine
vs. L-lysine-d- amphetamine following oral administration (60
mg/kg) Plasma Amphetamine Concentrations (ng/ml) L-lysine-d-
d-amphetamine amphetamine Hours Mean .+-.SD CV Mean .+-.SD CV 0 NA
NA NA NA NA NA 0.25 3,721 286 8 169 93 55 0.5 3,566 560 16 259 138
53 1 3,556 442 12 420 173 41 1.5 4,142 381 9 506 169 33 3 NA NA NA
686 222 32 5 NA NA NA 612 67 11 8 NA NA NA 870 NA NA
TABLE-US-00017 TABLE 13 Pharmacokinetic parameters of d-amphetamine
following oral administration of d-amphetamine or
L-lysine-d-amphetamine 12 mg/kg 30 mg/kg 60 mg/kg Parameter Amp
K-Amp Amp K-Amp Amp K-Amp AUG (ng h/ml) 2,738 1,958 12,623* 2,387*
5,081** 476** Percent 100 72 100 19 100 9 C.sub.max (ng/ml) 621 352
3,726 231 4,101 647 Percent 100 57 100 6 100 16 T.sub.max (hours)
0.938 3.5 NA NA NA NA Percent 100 373 NA NA NA NA *0-5 h **0-1.5
h
Example 8
Oral Bioavailability of L-lysine-d-amphetamine Dimesylate at
Various Doses Approximating a Range of Therapeutic Human Doses
Compared to a Suprapharmacological Dose
[0278] Male Sprague-Dawley rats were provided water ad libitum,
fasted overnight, and dosed by oral gavage with 1.5, 3, 6, 12, and
60 mg/kg of amphetamine sulfate or L-lysine-d-amphetamine
containing the equivalent amounts of d-amphetamine. Concentrations
of d-amphetamine were measured by ELISA.
[0279] It has been demonstrated that when lysine is conjugated to
the active agent d-amphetamine, the levels of d-amphetamine at 30
minutes post-administration are decreased by approximately 50% over
a dosage range of 1.5 to 12 mg/kg. However, when a
suprapharmacological dose (60 mg/kg) is given, the levels of
d-amphetamine from L-lysine-d-amphetamine only reached 8% of those
seen for d-amphetamine sulfate (See Table 14, Table 15, and FIG.
15). The substantial decrease in oral bioavailability at a high
dose greatly reduces the abuse potential of
L-lysine-d-amphetamine.
TABLE-US-00018 TABLE 14 Levels of d-amphetamine vs. dosage at 0.5 h
post dosing with d-amphetamine sulfate Dose mg/kg 1.5 3 6 12 60
ng/ml 0.5 h 109 .+-. 59 196 .+-. 72 294 .+-. 202 344 .+-. 126 323
.+-. 973 Percent 100 100 100 100 100
TABLE-US-00019 TABLE 15 Levels of d-amphetamine vs. dosage at 0.5 h
post dosing with L-lysine-d-amphetamine Dose mg/kg 1.5 3 6 12 60
ng/ml 0.5 h 45 .+-. 10 86 .+-. 26 129 .+-. 46 172 .+-. 113 266 .+-.
18 Percent 41 44 44 50 8
Example 9
Decreased Oral Bioavailability of L-lysine-d-amphetamine Dimesylate
at a High Dose
[0280] An additional oral PK study illustrated in FIG. 16 shows the
d-amphetamine blood levels of a 60 mg/kg dose over an 8 h time
course. In the case of d-amphetamine, blood levels quickly reached
a very high level, and 8 of 12 animals either died or were
sacrificed due to acute symptoms of toxicity. Blood levels (Table
16 and Table 17) of animals administered L-lysine-d-amphetamine, on
the other hand, did not peak until 5 hours and reached only a
fraction of the levels of the animals receiving amphetamine. (Note:
Valid data past 3 h for d-amphetamine could not be determined due
to death and sacrifice of animals).
TABLE-US-00020 TABLE 16 Mean plasma concentrations of d-amphetamine
vs. L-lysine- d-amphetamine following oral administration of a high
dose (60 mg/kg) Plasma Amphetamine Concentrations (ng/ml)
L-lysine-d- d-amphetamine amphetamine Hours Mean .+-.SD CV Mean
.+-.SD CV 0 NA NA NA NA NA NA 0.25 2174 907 42 35 17 48 0.5 2643
578 22 81 33 41 1 2828 1319 47 212 30 14 1.5 2973 863 29 200 79 40
3 2944* 95 3 440 133 30 5 153* NA NA 565 100 18 8 1309** NA NA 410
206 50 *n = 2 **n = 1
TABLE-US-00021 TABLE 17 Pharmacokinetic parameters of d-amphetamine
vs. L-lysine- d-amphetamine Mean AUC Percent C.sub.max Percent Peak
Percent Drug ng h/ml d-Amp (ng/ml) d-Amp (ng/ml) d-Amp d- 13420 100
3623 100 2973 100 amphetamine L-lysine-d- 3,143 39 582 16 565 19
amphetamine
Example 10
Oral Bioavailability of d-amphetamine Following Administration of
an Extended Release Formulation (Intact or Crushed) or
L-lysine-d-amphetamine Dimesylate
[0281] Doses of an extended release formulation of d-amphetamine
sulfate (Dexedrine Spansule.RTM. capsules, GlaxoSmithKline) were
orally administered to rats as intact capsules or as crushed
capsules and compared to a dose of L-lysine-d-amphetamine
containing an equivalent amount of d-amphetamine base (FIG. 20).
The crushed capsules showed an increase in C.sub.max and
AUC.sub.inf of 84 and 13 percent, respectively, as compared to
intact capsules (Table 18 and Table 19). In contrast, C.sub.max and
AUC.sub.inf of d-amphetamine following administration of
L-lysine-d-amphetamine were similar to that of the intact capsule
illustrating that extended release is inherent to the compound
itself and can not be circumvented by simple manipulation.
TABLE-US-00022 TABLE 18 Time-course concentrations of d-amphetamine
following oral administration of extended release Dexedrine
Spansule .RTM. capsules, crushed extended release Dexedrine
Spansule .RTM. capsules, or L-lysine-d-amphetamine (3 mg/kg d-
amphetamine base) Plasma Concentration (ng/ml) Crushed L-lysine-d-
Hours Intact Spansule .RTM. Capsule Spansule .RTM. Capsule
amphetamine 0 0 0 0 0.25 32 46 3 0.5 33 85 5 1 80 147 34 1.5 61 101
60 3 64 66 76 5 46 39 66 8 34 12 38
TABLE-US-00023 TABLE 19 Pharmacokinetic parameters of d-amphetamine
following oral administration of extended release Dexedrine
Spansule .RTM. capsules, crushed extended release Dexedrine
Spansule .RTM. capsules, or L-lysine-d-amphetamine (3 mg/kg
d-amphetamine base) Intact Crushed L-lysine-d- Parameter Spansule
.RTM. Capsule Spansule .RTM. Capsule amphetamine AUC.sub.0-8 h 399
449 434 (ng h/ml) Percent 100 113 109 C.sub.max (ng/ml) 80 147 76
Percent 100 184 95 T.sub.max (hours) 1 1 3 Percent 100 100 300
[0282] This example illustrates the advantage of the invention over
conventional controlled release formulations of d-amphetamine.
Example 11
Decreased Intranasal Bioavailability of L-lysine-d-amphetamine Vs.
Amphetamine
[0283] a. Intranasal (IN) Bioavailability of L-lysine-d-amphetamine
Hydrochloride
[0284] Male Sprague-Dawley rats were dosed by intranasal
administration with 3 mg/kg of amphetamine sulfate or
L-lysine-d-amphetamine hydrochloride containing the equivalent
amounts of d-amphetamine. L-lysine-d-amphetamine did not release
any significant amount of d-amphetamine into circulation by IN
administration. Mean (n=4) plasma amphetamine concentration curves
of amphetamine vs. L-lysine-d-amphetamine are shown in FIG. 17.
Pharmacokinetic parameters for IN administration of
L-lysine-d-amphetamine are summarized in Table 20.
TABLE-US-00024 TABLE 20 Pharmacokinetic parameters of d-amphetamine
vs. L-lysine-d-amphetamine hydrochloride by IN administration AUC
(0-1.5 h) Percent C.sub.max Percent Drug ng h/ml d-amphetamine
(ng/ml) d-amphetamine Amphetamine 727 100 1,377 100 L-lysine-d- 4
0.5 7 0.5 amphetamine
[0285] b. Intranasal Bioavailability of L-lysine-d-amphetamine
Dimesylate
[0286] The process of part a was repeated using
L-lysine-d-amphetamine mesylate salt:
TABLE-US-00025 TABLE 21 Pharmacokinetic parameters of d-amphetamine
vs. L-lysine-d-amphetamine mesylate salt by IN administration AUC
(0-1.0 h) Percent C.sub.max Percent Drug ng h/ml d-amphetamine
(ng/ml) d-amphetamine Amphetamine 573 100 1114 100 L-lysine-d- 25 4
26 2 amphetamine mesylate salt
[0287] This example illustrates that when lysine is conjugated to
the active agent d-amphetamine, the bioavailability by the
intranasal route is substantially decreased, thereby diminishing
the ability to abuse the drug by this route.
Example 12
Intravenous Bioavailability of Amphetamine Vs.
L-lysine-d-amphetamine Dimesylate
[0288] Male Sprague-Dawley rats were dosed by intravenous tail vein
injection with 1.5 mg/kg of d-amphetamine or L-lysine-d-amphetamine
containing the equivalent amount of amphetamine. As observed with
IN dosing, the conjugate did not release a significant amount of
d-amphetamine. Mean (n=4) plasma concentration curves of
amphetamine vs. L-lysine-d-amphetamine are shown in FIG. 19.
Pharmacokinetic parameters for IV administration of
L-lysine-d-amphetamine are summarized in Table 22.
TABLE-US-00026 TABLE 22 Pharmacokinetic parameters of d-amphetamine
vs. L-lysine-d-amphetamine by IV administration AUC (0-1.5 h)
Percent C.sub.max Percent Drug ng h/ml Amphetamine (ng/ml)
Amphetamine Amphetamine 190 100 169 100 K-amphetamine 6 3 5 3
[0289] This example illustrates that when lysine is conjugated to
the active agent amphetamine, the bioavailability of amphetamine by
the intravenous route is substantially decreased, thereby
diminishing the ability to abuse the drug by this route.
Example 13
Oral Bioavailability of L-lysine-d-amphetamine Dimesylate Compared
to d-Amphetamine at Escalating Doses
[0290] The fraction of intact L-lysine-d-amphetamine absorbed
following oral administration in rats increased non-linearly in
proportion to escalating doses from 1.5 to 12 mg/kg (FIG. 21-FIG.
25). The fraction absorbed at 1.5 mg/kg was only 2.6 percent
whereas it increased to 24.6 percent by 12 mg/kg. The fraction
absorbed fell to 9.3 percent at the high dose of 60 mg/kg.
T.sub.max ranged from 0.25 to 3 hours, and peak concentrations
occurred earlier for L-lysine-d-amphetamine than for d-amphetamine.
L-lysine-d-amphetamine was cleared more rapidly than d-amphetamine
with nearly undetectable concentrations by 8 hours at the lowest
dose.
[0291] The bioavailability (AUC) of d-amphetamine from each drug
administered was approximately equivalent at low doses. T.sub.max
for d-amphetamine from L-lysine-d-amphetamine ranged from 1.5 to 5
hours as compared to 0.5 to 1.5 following administration of
d-amphetamine sulfate. The difference in T.sub.max was greater at
higher doses. C.sub.max of d-amphetamine from
L-lysine-d-amphetamine was reduced by approximately half as
compared to the C.sub.max of d-amphetamine from d-amphetamine
sulfate administration at doses of 1.5 to 6 mg/kg, doses
approximating therapeutic human equivalent doses (HEDs). Thus, at
therapeutic doses, the pharmacokinetics of d-amphetamine from
L-lysine-d-amphetamine resembled that of a sustained release
formulation.
[0292] HEDs are defined as the equivalent dose for a 60 kg person
in accordance to the body surface area of the animal model. The
adjustment factor for rats is 6.2. The HED for a rat dose of 1.5
mg/kg of d-amphetamine, for example, is equivalent to
1.5/6.2.times.60=14.52 d-amphetamine base; which is equivalent to
14.52/0.7284=19.9 mg d-amphetamine sulfate, when adjusted for the
salt content.
TABLE-US-00027 TABLE 23 Human Equivalent Doses (HEDs) of
d-amphetamine sulfate Rat dose of d-amphetamine Human equivalent
dose (HED) of (mg/kg) d-amphetamine sulfate (mg) 1.5 19.9 3 39.9 6
79.7 12 159.4 30 399 60 797.2
[0293] At suprapharmacological doses (12 and 60 mg/kg), C.sub.max
was reduced by 73 and 84 percent, respectively, as compared to
d-amphetamine sulfate. For these high doses, the AUCs for
d-amphetamine from L-lysine-d-amphetamine were substantially
decreased compared to those of d-amphetamine sulfate, with the
AUC.sub.inf reduced by 76% at the highest dose (60 mg/kg). At 60
mg/kg, the levels of d-amphetamine from d-amphetamine sulfate
spiked rapidly; the experimental time course could not be completed
due to extreme hyperactivity necessitating humane euthanasia.
[0294] In summary, oral bioavailability of d-amphetamine from
L-lysine-d-amphetamine decreased to some degree at higher doses.
However, pharmacokinetics with respect to dose were nearly linear
for L-lysine-d-amphetamine at doses from 1.5 to 60 mg/kg with the
fraction absorbed ranging from 52 to 81 percent (extrapolated from
1.5 mg/kg dose). Pharmacokinetics of d-amphetamine sulfate was also
nearly linear at lower doses of 1.5 to 6 mg/kg with the fraction
absorbed ranging from 62 to 84 percent. In contrast to
L-lysine-d-amphetamine, however, parameters were disproportionately
increased at higher doses for d-amphetamine sulfate with the
fraction absorbed calculated as 101 and 223 percent (extrapolated
from 1.5 mg/kg dose), respectively, for the suprapharmacological
doses of 12 and 60 mg/kg.
[0295] The results suggest that the capacity for clearance of
d-amphetamine when delivered as the sulfate salt becomes saturated
at the higher doses whereas the gradual hydrolysis of
L-lysine-d-amphetamine precludes saturation of d-amphetamine
elimination at higher doses. The difference in proportionality of
dose to bioavailability (C.sub.max and AUC) for d-amphetamine and
L-lysine-d-amphetamine is illustrated in FIG. 26-FIG. 28. The
pharmacokinetic properties of L-lysine-d-amphetamine as compared to
d-amphetamine at the higher doses decrease the ability to escalate
doses. This improves the safety and reduces the abuse liability of
L-lysine-d-amphetamine as a method of delivering d-amphetamine for
the treatment of ADHD or other indicated conditions.
Example 14
Intranasal Bioavailability of L-lysine-d-amphetamine Dimesylate
Compared to d-amphetamine
[0296] As shown in FIG. 31 and FIG. 32, bioavailability of
d-amphetamine following bolus intranasal administration of
L-lysine-d-amphetamine was approximately 5 percent of that of the
equivalent d-amphetamine sulfate dose with AUC.sub.inf values of 56
and 1032, respectively. C.sub.max of d-amphetamine following
L-lysine-d-amphetamine administration by the intranasal route was
also about 5 percent of that of the equivalent amount of
d-amphetamine sulfate with values of 78.6 ng/mL and 1962.9 ng/mL,
respectively. T.sub.max of d-amphetamine concentration was delayed
substantially for L-lysine-d-amphetamine (60 minutes) as compared
to T.sub.max of d-amphetamine sulfate (5 minutes), reflecting the
gradual hydrolysis of L-lysine-d-amphetamine. Also, a high
concentration of intact L-lysine-d-amphetamine was detected
following intranasal administration. These results suggest that
intranasal administration of L-lysine-d-amphetamine provides only
minimal hydrolysis of L-lysine-d-amphetamine and thus only minimal
release of d-amphetamine.
Example 15
Intravenous Bioavailability of L-lysine-d-amphetamine Dimesylate
Compared to d-amphetamine
[0297] As shown in FIG. 33 and FIG. 34, bioavailability of
d-amphetamine following bolus intravenous administration of
L-lysine-d-amphetamine was approximately one-half that of the
equivalent d-amphetamine sulfate dose with AUC.sub.inf values of
237.8 and 420.2, respectively. C.sub.max of d-amphetamine following
L-lysine-d-amphetamine administration was only about one-fourth
that of the equivalent amount of d-amphetamine with values of 99.5
and 420.2, respectively. T.sub.max of d-amphetamine concentration
was delayed substantially for L-lysine-d-amphetamine (30 minutes)
as compared to T.sub.max of d-amphetamine sulfate (5 minutes),
reflecting the gradual hydrolysis of L-lysine-d-amphetamine. In
conclusion, the bioavailability of d-amphetamine by the intravenous
route is substantially decreased and delayed when given as
L-lysine-d-amphetamine. Moreover, bioavailability is less than that
obtained by oral administration of the equivalent dose of
L-lysine-d-amphetamine.
Summary of LC/MS/MS Bioavailability Data in Rats
[0298] The following tables summarize the bioavailability data
collected in the experiments discussed in Examples 13-15. Table 24,
Table 25, and Table 26 summarize the pharmacokinetic parameters of
d-amphetamine following oral, intranasal, and intravenous
administration, respectively, of d-amphetamine or
L-lysine-d-amphetamine.
TABLE-US-00028 TABLE 24 Pharmacokinetic parameters of d-amphetamine
following oral administration of L-lysine-d-amphetamine or
d-amphetamine at escalating doses AUC.sub.inf Dose C.sub.max
T.sub.max AUC.sub.0-8 AUC.sub.inf alt. F AUC/Dose C.sub.max/Dose
Drug (mg/kg) (ng/mL) (h) (ng h/mL) (ng h/mL) calc.* (%) (ng h
kg/mL/mg) (ng kg/mL/mg) K-Amp 1.5 59.6 3 308 331 376 61 220.7 39.7
Amp 1.5 142.2 0.5 446 461 483 84 307.3 94.8 K-Amp 3 126.9 1.5 721
784 963 72 261.3 42.3 Amp 3 217.2 1.5 885 921 1,059 84 307.0 72.4
K-Amp 6 310.8 3 1,680 1,797 2,009 82 299.5 51.8 Amp 6 815.3 0.25
1,319 1,362 1429 62 227.0 135.9 K-Amp 12 412.6 5 2,426 2,701 2,701
62 225.1 34.4 Amp 12 1,533.1 0.25 4,252 4,428 4,636 101 369.0 127.8
K-Amp 60 2,164.3 5 9995.1 11,478 11,478 52 191.3 36.1 Amp 60 13,735
1 14,281** 48,707 48,707 223 811.8 228.9 *An alternative
calculation of AUC.sub.inf can be performed using WinNonlin .RTM.
software (Version 4.1, Pharsight, Inc., Mountain View, California).
**AUC (0-1.5)
TABLE-US-00029 TABLE 25 Pharmacokinetic parameters of d-amphetamine
following bolus intravenous administration of
L-lysine-d-amphetamine (1.5 mg/kg) Dose C.sub.max T.sub.max
AUC.sub.0-24 AUC.sub.0-24 AUC.sub.inf AUC.sub.inf Route Drug
(mg/kg) (ng/mL) (h) (ng h/mL) alt. calc.* (ng h/mL) alt. calc.* IV
K-Amp 1.5 99.5 0.5 237.8 207 237.9 218 IV Amp 1.5 420.2 0.083 546.7
511 546.9 521
TABLE-US-00030 TABLE 26 Pharmacokinetic parameters of d-amphetamine
following intranasal administration of L-lysine-d-amphetamine Dose
C.sub.max T.sub.max AUC.sub.0-1 AUC.sub.inf AUC.sub.inf Route Drug
(mg/kg) (ng/mL) (h) (ng h/mL) (ng h/mL) alt. calc.* IN K-Amp 3 78.6
1 56 91 NA IN Amp 3 1962.9 0.083 1032 7291 1,267
[0299] Table 27, Table 28, and Table 29 summarize the
pharmacokinetic parameters of L-lysine-d-amphetamine following
oral, intravenous, and intranasal administration of
L-lysine-d-amphetamine.
TABLE-US-00031 TABLE 27 Pharmacokinetic parameters of
L-lysine-d-amphetamine following oral administration of
L-lysine-d-amphetamine at escalating doses Dose C.sub.max T.sub.max
AUC.sub.0-8 AUC.sub.inf AUC.sub.inf F Route Drug (mg/kg) (ng/ml)
(ng/ml) (ng h/mL) (ng h/mL) alt. calc.* (%) Oral K-Amp 1.5 36.5
0.25 59.4 60 60 2.6 Oral K-Amp 3 135.4 1.5 329.7 332.1 331 7.2 Oral
K-Amp 6 676.8 0.25 1156.8 1170.8 1,176 12.8 Oral K-Amp 12 855.9 1
4238.6 4510.4 5,169 24.6 Oral K-Amp 60 1870.3 3 8234.3 8499.9 8,460
9.3
TABLE-US-00032 TABLE 28 Pharmacokinetic parameters of
L-lysine-d-amphetamine following bolus intravenous administration
of L-lysine-d-amphetamine Dose C.sub.max T.sub.max AUC.sub.0-24
AUC.sub.inf Route Drug (mg/kg) (ng/mL) (h) (ng h/mL) (ng h/mL) IV
K-Amp 1.5 4513.1 0.083 2,282 2,293
TABLE-US-00033 TABLE 29 Pharmacokinetic parameters of
L-lysine-d-amphetamine following intranasal administration of
L-lysine-d-amphetamine Dose C.sub.max T.sub.max AUC.sub.0-1
AUC.sub.inf Route Drug (mg/kg) (ng/mL) (h) (ng h/mL) (ng h/mL) IN
K-Amp 3 3345.1 0.25 2,580 9,139
[0300] Table 30 and Table 31 summarize the percent bioavailability
of d-amphetamine following oral, intranasal, and intravenous
administration of L-lysine-d-amphetamine as compared to
d-amphetamine sulfate.
TABLE-US-00034 TABLE 30 Percent bioavailability (AUC.sub.inf) of
d-amphetamine following administration of L-lysine-d-amphetamine by
various routes as compared to bioavailability following
administration of d-amphetamine sulfate Dose (mg/kg) d-amphetamine
base 1.5 3 6 12 60 HED 19.9 39.9 79.7 159.4 797.2 Oral 72 85 132 61
24 IV 43 NA NA NA NA IN NA 1 NA NA NA
TABLE-US-00035 TABLE 31 Percent bioavailability (C.sub.max) of
d-amphetamine following administration of L-lysine-d-amphetamine by
various routes as compared to bioavailability following
administration of d-amphetamine sulfate Dose (mg/kg) d-amphetamine
base 1.5 3 6 12 60 HED 19.9 39.9 79.7 159.4 797.2 Oral 42 58 38 27
16 IV 24 NA NA NA NA IN NA 4 NA NA NA
[0301] Table 32-Table 37 summarize the time-course concentrations
of d-amphetamine and L-lysine-d-amphetamine following oral,
intranasal, and intravenous administration of d-amphetamine or
L-lysine-d-amphetamine.
TABLE-US-00036 TABLE 32 Time-course concentrations of d-amphetamine
following bolus intravenous administration of
L-lysine-d-amphetamine or d-amphetamine sulfate (1.5 mg/kg) Time
Concentration (ng/ml) (hours) K-Amp Amp sulfate 0 0 0 0.083 52.8
420.2 0.5 99.5 249.5 1.5 47.1 97.9 3 21.0 38.3 5 9.0 13.2 8 3.7 4.3
24 0.1 0.2
TABLE-US-00037 TABLE 33 Time-course concentrations of
L-lysine-d-amphetamine following bolus intravenous administration
of L-lysine-d-amphetamine (1.5 mg/kg) K-Amp Time concentration
(hours) (ng/ml) 0 0 0.083 4513.1 0.5 1038.7 1.5 131.4 3 19.3 5 17.9
8 8.7 24 11.5
TABLE-US-00038 TABLE 34 Time-course concentrations of d-amphetamine
following oral administration of L-lysine-d-amphetamine at various
doses Time Concentration (ng/ml) (hours) 1.5 mg/kg 3 mg/kg 6 mg/kg
12 mg/kg 60 mg/kg 0 0 0 0 0 0 0.25 20.5 25.3 96 54.3 90.9 0.5 34
40.9 140.2 96 175.1 1 46.7 95.1 225.9 233.3 418.8 1.5 40.7 126.9
268.4 266 440.7 3 59.6 105 310.8 356.8 1145.5 5 38.6 107.6 219.5
412.6 2164.3 8 17.1 48 86 225.1 1227.5
TABLE-US-00039 TABLE 35 Time-course concentrations of d-amphetamine
following oral administration of d-amphetamine sulfate at various
doses Time Concentration (ng/ml) (hours) 1.5 mg/kg 3 mg/kg 6 mg/kg
12 mg/kg 60 mg/kg 0 0 0 0 0 0 0.25 107.1 152.6 815.3 1533.1 6243.6
0.5 142.2 198.4 462.7 1216 7931.6 1 105.7 191.3 301.3 828.8 13735.2
1.5 129.5 217.2 314 904.8 11514.9 3 52.6 135.3 134.6 519.9 NA 5
29.5 73.5 77.4 404.3 NA 8 11.5 25.7 31.8 115.4 NA
TABLE-US-00040 TABLE 36 Time-course concentrations of d-amphetamine
following intranasal administration of L-lysine-d-amphetamine or
d-amphetamine sulfate (3 mg/kg) Time Concentration (ng/ml) (hours)
K-Amp Amp sulfate 0 0 0 0.083 31.2 1962.9 0.25 45.3 1497.3 0.5 61.3
996.2 1 78.6 404.6 AUC 56 1032.3
TABLE-US-00041 TABLE 37 Time-course concentrations of
L-lysine-d-amphetamine following intranasal administration of
L-lysine-d-amphetamine (3 mg/kg) Concentration Time (ng/ml) (hours)
K-Amp 0 0 0.083 3345.1 0.25 3369.7 0.5 2985.8 1 1359.3
Example 16
Bioavailability of L-lysine-d-amphetamine Dimesylate or
d-amphetamine Sulfate in Dogs (LC/MS/MS Analysis)
[0302] Example Experimental Design:
[0303] This was a non-randomized, two-treatment crossover study.
All animals were maintained on their normal diet and were fasted
overnight prior to each dose administration. L-lysine-d-amphetamine
dose was based on the body weight measured on the morning of each
dosing day. The actual dose delivered was based on syringe weight
before and after dosing. Serial blood samples were obtained from
each animal by direct venipuncture of a jugular vein using
vacutainer tubes containing sodium heparin as the anticoagulant.
Derived plasma samples were stored frozen until shipment to Quest
Pharmaceutical Services, Inc. (Newark, Del.). Pharmacokinetic
analysis of the plasma assay results was conducted by Calvert.
Animals were treated as follows:
TABLE-US-00042 Number Dose Dose of Dogs/ Route of Conc. Dose Vol.
Level Sex Administration Treatment (mg/mL) (mL/kg) (mg/kg) 3/M PO 1
0.2 10 1 3/M IV 2 1 2 1
[0304] Administration of the Test Article:
[0305] Oral: The test article was administered to each animal via a
single oral gavage. On Day 1, animals received the oral dose by
gavage using an esophageal tube attached to a syringe. Dosing tubes
were flushed with approximately 20 mL tap water to ensure the
required dosing solution was delivered.
[0306] Intravenous: On Day 8, animals received
L-lysine-d-amphetamine as a single 30-minute intravenous infusion
into a cephalic vein.
[0307] Sample Collection:
[0308] Dosing Formulations: Post-dosing, remaining dosing
formulation was saved and stored frozen.
[0309] Blood: Serial blood samples (2 mL) were collected using
venipuncture tubes containing sodium heparin. Blood samples were
taken at 0, 0.25, 0.5, 1, 2, 4, 8, 12, 24, 48, and 72 hours
post-oral dosing. Blood samples were collected at 0, 0.167, 0.33,
0.49 (prior to stop of infusion), 0.583, 0.667, 0.75, 1, 2, 3, 4,
8, 12, and 23 hours post-intravenous infusion start. Collected
blood samples were chilled immediately.
[0310] Plasma: Plasma samples were obtained by centrifugation of
blood samples. Duplicate plasma samples (about 0.2 mL each) were
transferred into prelabeled plastic vials and stored frozen at
approximately -70.degree. C.
[0311] Sample Assay:
[0312] Plasma samples were analyzed for L-lysine-d-amphetamine and
d-amphetamine using a validated LC-MS/MS method with an LLOQ of 1
ng/mL for both analytes.
[0313] Microsoft Excel (Version 6, Microsoft Corp., Redmond, Wash.)
was used for calculation of mean plasma concentration and graphing
of the plasma concentration-time data. Pharmacokinetic analysis
(non-compartmental) was performed using the WinNonlin.RTM. software
program (Version 4.1, Pharsight, Inc. Mountain View, Calif.). The
maximum concentration (C.sub.max) and the time to
C.sub.max(T.sub.max) were observed values. The area under the
plasma concentration-time curve (AUC) was determined using
linear-log trapezoidal rules. The apparent terminal rate constant
(.lamda.z) was derived using linear least-squares regression with
visual inspection of the data to determine the appropriate number
of points (minimum of 3 data points) for calculating .lamda.z. The
AUC.sub.0-inf was calculated as the sum of AUC.sub.0-t and
Cpred/.lamda.z, where Cpred was the predicted concentration at the
time of the last quantifiable concentration. The plasma clearance
(CL/F) was determined as the ratio of Dose/AUC.sub.0-inf. The mean
residence time (MRT) was calculated as the ratio of
AUMC.sub.0-inf/AUC.sub.0-inf, where AUMC.sub.0-inf was the area
under the first moment curve from the time zero to infinity. The
volume of distribution at steady state (V.sub.ss) was estimated as
CL*MRT. Half-life was calculated as ln 2/.lamda.z. The oral
bioavailability (F) was calculated as the ratio of AUC.sub.0-inf
following oral dosing to AUC.sub.0-inf following intravenous
dosing. Descriptive statistics (mean and standard deviation) of the
pharmacokinetic parameters were calculated using Microsoft
Excel.
[0314] The objectives of this study were to characterize the
pharmacokinetics of L-lysine-d-amphetamine and d-amphetamine
following administration of L-lysine-d-amphetamine in male beagle
dogs. As shown in FIG. 35, in a cross-over design,
L-lysine-d-amphetamine was administered to 3 male beagle dogs
orally and intravenously. Blood samples were collected up to 24 and
72 hours after the intravenous and oral doses, respectively.
[0315] The mean L-lysine-d-amphetamine and d-amphetamine plasma
concentration-time profiles following an intravenous or oral dose
of L-lysine-d-amphetamine are presented in FIG. 37 and FIG. 38,
respectively. Comparative profiles of L-lysine-d-amphetamine to
d-amphetamine following both routes are depicted in FIG. 35 and
FIG. 36. Individual plots are depicted in FIG. 39 and FIG. 40. The
pharmacokinetic parameters are summarized in Table 38-Table 46. The
results indicate that administration of L-lysine-d-amphetamine, as
compared to administration of amphetamine, leads to a decrease in
patient to patient variability of amphetamine levels (see
respective CV values in Table 45).
[0316] Following a 30-minute intravenous infusion of
L-lysine-d-amphetamine, the plasma concentration reached a peak at
the end of the infusion. Post-infusion L-lysine-d-amphetamine
concentration declined very rapidly in a biexponential manner, and
fell below the quantifiable limit (1 ng/mL) by approximately 8
hours post-dose. Results of non-compartmental pharmacokinetic
analysis indicate that L-lysine-d-amphetamine is a high clearance
compound with a moderate volume of distribution (V.sub.ss)
approximating total body water (0.7 L/kg). The mean clearance value
was 2087 mL/hkg (34.8 mL/minkg) and was similar to the hepatic
blood flow in the dog (40 mL/minkg).
[0317] L-lysine-d-amphetamine was rapidly absorbed after oral
administration with T.sub.max at 0.5 hours in all three dogs. Mean
absolute oral bioavailability was 33%, which suggests that
L-lysine-d-amphetamine is very well absorbed in the dog. The
apparent terminal half-life was 0.39 hours, indicating rapid
elimination, as observed following intravenous administration.
[0318] Plasma concentration-time profiles of d-amphetamine
following intravenous or oral administration of
L-lysine-d-amphetamine were similar. See Table 39. At a 1 mg/kg
oral dose of L-lysine-d-amphetamine, the mean C.sub.max of
d-amphetamine was 104.3 ng/mL. The half-life of d-amphetamine was
3.1 to 3.5 hours, much longer when compared to
L-lysine-d-amphetamine.
[0319] In this study, L-lysine-d-amphetamine was infused over a 30
minute time period. Due to rapid clearance of
L-lysine-d-amphetamine it is likely that bioavailability of
d-amphetamine from L-lysine-d-amphetamine would decrease if a
similar dose were given by intravenous bolus injection. Even when
given as an infusion the bioavailability of d-amphetamine from
L-lysine-d-amphetamine did not exceed that of a similar dose given
orally and the time to peak concentration was substantially
delayed. This data further supports that L-lysine-d-amphetamine
affords a decrease in the abuse liability of d-amphetamine by
intravenous injection.
TABLE-US-00043 TABLE 38 Pharmacokinetic parameters of
L-lysine-d-amphetamine in male beagle dogs following oral or
intravenous administration of L-lysine-d-amphetamine (1 mg/kg
d-amphetamine base) Dose C.sub.max AUC.sub.inf CL/F V.sub.ss Route
mg/kg ng/mL T.sub.max.sup.a h ng h/mL t.sub.1/2 h MRT h mL/h kg
mL/kg F % IV 1 1650 0.49 964 0.88 0.33 2087 689 NA (0.00) (178)
(0.49-0.49) (97.1) (0.2) (0.03) (199) (105.9) Oral 1 328.2 0.5 319
0.39 0.81 6351 NA 33 (0.00) (91.9) (0.5-0.5) (46.3) (0.1) (0.19)
(898.3) (1.9) .sup.amedian (range) Abbreviations of pharmacokinetic
parameters are as follows: C.sub.max, maximum observed plasma
concentration; T.sub.max, time when C.sub.max observed;
AUC.sub.0-t, total area under the plasma concentration versus time
curve from 0 to the last data point; AUC.sub.0-inf, total area
under the plasma concentration versus time curve; t.sub.1/2,
apparent terminal half-life; MRT, mean residence time; CL/F, oral
clearance; V.sub.ss, volume of distribution at steady state; F,
bioavailability.
TABLE-US-00044 TABLE 39 Pharmacokinetic parameters of d-amphetamine
in male beagle dogs following oral or intravenous administration of
L-lysine-d- amphetamine (1 mg/kg d-amphetamine base) Dose C.sub.max
T.sub.max.sup.a AUC.sub.inf t.sub.1/2 Route (mg/kg) (ng/mL) (h) (ng
h/mL) (h) IV 1 113.2 1.0 672.5 3.14 (0.00) (3.2) (0.67-2.0) (85.7)
(0.4) Oral 1 104.3 2.0 728.0 3.48 (0.00) (21.8) (2-2) (204.9) (0.4)
.sup.amedian (range)
TABLE-US-00045 TABLE 40 Pharmacokinetics of L-lysine-d-amphetamine
in male beagle dogs following 30 min intravenous administration of
L-lysine-d-amphetamine (1 mg/kg d- amphetamine base) C.sub.max
T.sub.max.sup.a AUC.sub.0-t AUC.sub.inf t.sub.1/2 CL V.sub.ss MRT
Dog ID (ng/mL) (h) (ng h/mL) (ng h/mL) (h) (mL/h/kg) (mL/kg) (h) 1
1470.3 0.49 898.2 900.2 0.72 2222 807.4 0.36 2 1826.4 0.49 1072.3
1076.1 ND.sup.b 1859 603.4 0.32 3 1654.2 0.49 914.1 916.9 1.05 2181
656.0 0.30 Mean 1650 0.49 961.5 964.4 0.88 2087 689.0 0.33 SD 178
0.49-0.49 96.0 97.1 0.2 199 105.9 0.03 .sup.amedian (range);
.sup.bnot determined CL, clearance following IV administration
TABLE-US-00046 TABLE 41 Pharmacokinetic parameters of
L-lysine-d-amphetamine in male beagle dogs following oral
administration of L-lysine-d-amphetamine (1 mg/kg d- amphetamine
base) C.sub.max T.sub.max.sup.a AUC.sub.0-t AUC.sub.inf t.sub.1/2
CL/F MRT F Dog ID (ng/mL) (h) (ng h/mL) (ng h/mL) (h) (mL/h/kg) (h)
(%) 1 350.2 0.5 275.3 277.1 0.24 7218 0.68 30.8 2 407.2 0.5 367.8
368.7 0.48 5424 0.74 34.3 3 227.4 0.5 310.8 312.0 0.45 6410 1.03
34.0 Mean 328.2 0.5 318.0 319.3 0.39 6351 0.81 33.0 SD 91.9 0.0
46.7 46.3 0.1 898.3 0.19 1.9 .sup.amedian (range)
TABLE-US-00047 TABLE 42 Pharmacokinetics of d-amphetamine in male
beagle dogs following 30 min. intravenous administration of
L-lysine-d-amphetamine (1 mg/kg d-amphetamine base) C.sub.max
T.sub.max.sup.a AUC.sub.0-t AUC.sub.inf t.sub.1/2 Dog ID (ng/mL)
(h) (ng h/mL) (ng h/mL) (h) 1 111.2 2.0 751.9 757.6 3.35 2 116.8
0.67 668.5 673.7 3.43 3 111.4 1.0 557.8 586.1 2.65 Mean 113.2 1.00
659.4 672.5 3.14 SD 3.2 0.67-2.0 97 85.7 0.4 .sup.amedian
(range)
TABLE-US-00048 TABLE 43 Pharmacokinetics of d-amphetamine in male
beagle dogs following oral administration of L-lysine-d-amphetamine
(1 mg/kg d-amphetamine base) C.sub.max T.sub.max.sup.a AUC.sub.0-t
AUC.sub.inf t.sub.1/2 Dog ID (ng/mL) (h) (ng h/mL) (ng h/mL) (h) 1
102.1 2.0 686.34 696.89 3.93 2 127.2 2.0 937.57 946.62 3.44 3 83.7
2.0 494.61 540.38 3.06 Mean 104.3 2.0 706.2 728.0 3.48 SD 21.8
2.0-2.0 222.1 204.9 0.4 .sup.amedian (range)
TABLE-US-00049 TABLE 44 Pharmacokinetics of d-amphetamine in male
beagle dogs following oral administration of L-lysine-d-amphetamine
or d-amphetamine sulfate (1.8 mg/kg d-amphetamine base) Mean Plasma
Standard Coefficient of Time Concentration Deviation (SD) Variation
(CV) (hours) Amp K-Amp Amp K-Amp Amp K-Amp 0 0 0 0 0 0 0 1 431.4
223.7 140.7 95.9 32.6 42.9 2 360 291.8 87.6 93.6 24.3 32.1 4 277.7
247.5 68.1 66 24.5 26.7 6 224.1 214.7 59.3 62.1 26.5 28.9 8 175.4
150 66.7 40.1 38.0 26.7 12 81.4 47.6 58.7 19 72.1 39.9 16 33 19.6
28.1 9 85.2 45.9 24 7.2 4.5 4.5 1.7 62.5 37.8
TABLE-US-00050 TABLE 45 Pharmacokinetics of d-amphetamine in female
beagle dogs following oral administration of L-lysine-d-amphetamine
or d-amphetamine sulfate (1.8 mg/kg d-amphetamine base) Mean Plasma
Standard Coefficient of Time Concentration Deviation (SD) Variation
(CV) (hours) Amp K-Amp Amp K-Amp Amp K-Amp 0 0 0 0 0 0 0 1 217.8
308.8 141.7 40.7 65.1 13.2 2 273.5 308 113.7 29.6 41.6 9.6 4 266
260.9 132.7 37.3 49.9 14.3 6 204.7 212.1 84.5 38.7 41.3 18.2 8
160.1 164.3 72.7 43.5 45.4 26.5 12 79.4 68.7 41.3 31 52.0 45.1 16
25.5 22.3 13.4 4.7 52.5 21.1 24 5.6 5.4 4.1 1.9 73.2 35.2
TABLE-US-00051 TABLE 46 Pharmacokinetic parameters of d-amphetamine
in male and female beagle dogs following oral administration of
L-lysine-d-amphetamine or d-amphetamine sulfate (1.8 mg/kg
d-amphetamine base) Males Females Compound Compound Parameter Amp
K-Amp Amp K-Amp AUC.sub.inf 3088.9 2382.2 2664.5 2569.9 Percent 100
77 100 96 C.sub.max 431.4 291.8 308.8 273.5 Percent 100 67 100 89
T.sub.max (hours) 1 2 1 2 Percent 100 200 100 200
Example 17
Delayed Cardiovascular Effects of L-lysine-d-amphetamine Dimesylate
as Compared to d-amphetamine Following Intravenous Infusion
[0320] Systolic and diastolic blood pressure (BP) are increased by
d-amphetamine even at therapeutic doses. Since
L-lysine-d-amphetamine is expected to release d-amphetamine (albeit
slowly) as a result of systemic metabolism, a preliminary study was
done using equimolar doses of d-amphetamine or
L-lysine-d-amphetamine to 4 dogs (2 male and 2 female). The results
suggest that the amide prodrug is inactive and that slow release of
some d-amphetamine, occurs beginning 20 minutes after the first
dose. Relative to d-amphetamine, however, the effects are less
robust. For example, the mean blood pressure is graphed in FIG. 43.
Consistent with previously published data (Kohli and Goldberg,
1982), small doses of d-amphetamine were observed to have rapid
effects on blood pressure. The lowest dose (0.202 mg/kg, equimolar
to 0.5 mg/kg of L-lysine-d-amphetamine) produced an acute doubling
of the mean BP followed by a slow recovery over 30 minutes.
[0321] By contrast, L-lysine-d-amphetamine produced very little
change in mean BP until approximately 30 minutes after injection.
At that time, pressure increased by about 20-50%. Continuous
release of d-amphetamine is probably responsible for the slow and
steady increase in blood pressure over the remaining course of the
experiment. Upon subsequent injections, d-amphetamine is seen to
repeat its effect in a non-dose dependent fashion. That is,
increasing dose 10-fold from the first injection produced a rise to
the same maximum pressure. This may reflect the state of
catecholamine levels in nerve terminals upon successive stimulation
of d-amphetamine bolus injections. Note that the rise in mean blood
pressure seen after successive doses of L-lysine-d-amphetamine
(FIG. 43) produces a more gradual and less intense effect. Similar
results were observed for left ventricular pressure (FIG. 44).
These results further substantiate the significant decrease in
d-amphetamine bioavailability by the intravenous route when given
as L-lysine-d-amphetamine. As a result the rapid onset of the
pharmacological effect of d-amphetamine that is sought by persons
injecting the drug is eliminated.
TABLE-US-00052 TABLE 47 Effects of L-lysine-d-amphetamine on
cardiovascular parameters in the anesthetized dog (mean values, n =
2) % % % % Treatment Time SAP Change DAP Change MAP Change LVP
Change 0.9% Saline 0 81 0 48 0 61 0 87 0 1 ml/kg 30 87 7 54 11 67
10 87 0 K-Amp 0 84 0 51 0 64 0 86 0 0.5 mg/kg 5 87 4 52 3 66 3 87 2
15 93 11 51 1 67 5 95 11 25 104 25 55 8 73 15 105 22 30 107 28 58
14 77 21 108 26 K-Amp 0 105 0 55 0 74 0 108 0 1.0 mg/kg 5 121 15 63
15 85 15 120 11 15 142 35 73 33 100 35 140 29 25 163 55 97 75 124
68 162 50 30 134 28 73 32 98 32 144 33 K-Amp 0 132 0 71 0 95 0 144
0 5.0 mg/kg 5 142 7 71 0 99 4 151 5 15 176 33 98 39 130 37 184 28
25 126 -5 69 -3 96 1 160 11 30 132 0 70 -1 99 4 163 13 SAP:
systolic arterial pressure (mmHg); MAP: mean arterial pressure
(mmHg); DAP: diastolic arterial pressure (mmHg); LVP: left
ventricular pressure (mmHg); % Change: percent change from
respective Time 0.
TABLE-US-00053 TABLE 48 Effects of d-amphetamine on cardiovascular
parameters in the anesthetized dog (mean values, n = 2) % % % %
Treatment Time SAP Change DAP Change MAP Change LVP Change 0.9%
Saline 0 110 0 67 0 84 0 105 0 1 ml/kg 30 108 -2 65 -3 82 -2 101 -3
d-amphetamine 0 111 0 67 0 84 0 104 0 0.202 mg/kg 5 218 97 145 117
176 109 214 107 15 168 52 97 45 125 49 157 52 25 148 34 87 30 110
31 142 37 30 140 26 80 20 103 23 135 30 d-amphetamine 0 139 0 78 0
101 0 133 0 0.404 mg/kg 5 240 73 147 88 187 85 238 79 15 193 39 112
44 145 43 191 43 25 166 19 92 17 122 20 168 26 30 160 16 87 11 117
16 163 22 d-amphetamine 0 158 0 87 0 115 0 162 0 2.02 mg/kg 5 228
44 128 48 169 47 227 40 15 196 24 107 23 142 23 200 24 25 189 20
102 17 135 17 192 19 30 183 16 98 13 129 12 187 16
Example 18
Pharmacodynamic (Locomotor) Response to Amphetamine Vs.
L-lysine-d-amphetamine diHCl by Oral Administration
[0322] Male Sprague-Dawley rats were provided water ad libitum,
fasted overnight, and dosed by oral gavage with 6 mg/kg of
amphetamine or L-lysine-d-amphetamine containing the equivalent
amount of d-amphetamine. Horizontal locomotor activity (HLA) was
recorded during the light cycle using photocell activity chambers
(San Diego Instruments). Total counts were recorded every 12
minutes for the duration of the test. Rats were monitored in three
separate experiments for 5, 8, and 12 hours, respectively. Time vs.
HLA counts for d-amphetamine vs. L-lysine-d-amphetamine is shown in
FIG. 45 and FIG. 46. In each experiment the time until peak
activity was delayed, and the pharmacodynamic effect was evident
for an extended period of time for L-lysine-d-amphetamine as
compared to d-amphetamine. The total activity counts for HLA of
Lys-Amp dosed rats were increased (11-41%) over those induced by
d-amphetamine in all three experiments.
TABLE-US-00054 TABLE 49 Locomotor activity of rats orally
administered d-amphetamine vs. L-lysine-d-amphetamine (5 h) Peak of
Time of Time of Total Total Activity activity Peak Last Count Test
Activity Counts Above (Counts (Counts Above 200 Material Counts
Baseline per 0.2 h) per 0.2 h) per 0.2 h Vehicle 4689 4174 80 1.4
-- K-Amp 6417 5902 318 1.8 5 h Amp 515 0 291 0.6 2.6 h
TABLE-US-00055 TABLE 50 Locomotor activity of rats orally
administered d-amphetamine vs. L-lysine-d-amphetamine (12 h) Peak
of Time of Time of Total Total Activity activity Peak Last Count
Test Activity Counts Above (Counts (Counts Above 200 Material
Counts Baseline per 0.2 h) per 0.2 h) per 0.2 h Vehicle 936 0 81
7.2 -- K-Amp 8423 7487 256 1.8 8.6 h Amp 6622 5686 223 0.6 6.4
h
Example 19
Pharmacodynamic Response to d-amphetamine Vs.
L-lysine-d-amphetamine diHCl by Intranasal Administration
[0323] Male Sprague-Dawley rats were dosed by intranasal
administration with d-amphetamine or L-lysine-d-amphetamine (1.0
mg/kg). In a second set of similarly dosed animals, carboxymethyl
cellulose (CMC) was added to the drug solutions at a concentration
of 62.6 mg/ml (approximately 2-fold higher than the concentration
of L-lysine-d-amphetamine and 5-fold higher than the d-amphetamine
content). The CMC drug mixtures were suspended thoroughly before
each dose was delivered. Locomotor activity was monitored using the
procedure described in Example 18. As shown in FIG. 47 and FIG. 48,
the activity vs. time (1 hour or 2 hours) is shown for
amphetamine/CMC vs. L-lysine-d-amphetamine and compared to that of
amphetamine vs. L-lysine-d-amphetamine CMC. As seen in FIG. 47,
addition of CMC to L-lysine-d-amphetamine decreased the activity
response of IN dosed rats to levels similar to the water/CMC
control, whereas no effect was seen on amphetamine activity by the
addition of CMC. The increase in activity over baseline of
L-lysine-d-amphetamine with CMC was only 9% compared to 34% for
L-lysine-d-amphetamine without CMC when compared to activity
observed for d-amphetamine dosed animals (Table 51). CMC had no
observable effect on d-amphetamine activity induced by IN
administration.
TABLE-US-00056 TABLE 51 Locomotor activity of intranasal
d-amphetamine vs. L-lysine-d-amphetamine with and without CMC Total
Activity Total Activity Counts Percent Drug n Counts (1 h) Above
Baseline Amp Amp 3 858 686 100 Amp CMC 3 829 657 100 K-Amp 4 408
237 35 K-Amp CMC 4 232 60 9 Water 1 172 0 0 Water CMC 1 172 0 0
Example 20
Pharmacodynamic Response to d-amphetamine Vs.
L-lysine-d-amphetamine diHCl by Intravenous Administration
[0324] Male Sprague-Dawley rats were dosed by intravenous
administration with d-amphetamine or L-lysine-d-amphetamine (1.0
mg/kg). The activity expressed as total activity counts over a
three hour period of time is shown in FIG. 49. The activity induced
by L-lysine-d-amphetamine was substantially decreased, and time to
peak activity was delayed. The increase in activity over baseline
of L-lysine-d-amphetamine was 34% for L-lysine-d-amphetamine when
compared to activity observed for d-amphetamine dosed animals
(Table 52).
TABLE-US-00057 TABLE 52 Total activity counts after intravenous
administration of d-amphetamine vs. L-lysine-d-amphetamine Total
Activity Percent Drug n Counts (3 h) Above Baseline Amp Amp 3 1659
1355 100 K-Amp 4 767 463 34 Water 1 304 0 0
Example 21
Decrease in Toxicity of Orally Administered L-lysine-d-amphetamine
diHCl
[0325] Three male and three female Sprague Dawley rats per group
were given a single oral administration of L-lysine-d-amphetamine
at 0.1, 1.0, 10, 60, 100, or 1000 mg/kg (Table 53). Each animal was
observed for signs of toxicity and death on Days 1-7 (with Day 1
being the day of the dose), and one rat/sex/group was necropsied
upon death (scheduled or unscheduled).
TABLE-US-00058 TABLE 53 Dosing chart for oral administration of
L-lysine-d-amphetamine toxicity testing No. of Animals Dose
Concentration Groups M F Test Article (mg/kg) (mg/mL) 1 3 3
L-lysine-d-amphetamine 0.1 0.01 2 3 3 L-lysine-d-amphetamine 1.0
0.1 3 3 3 L-lysine-d-amphetamine 10 1.0 4 3 3
L-lysine-d-amphetamine 60 6.0 5 3 3 L-lysine-d-amphetamine 100 10 6
3 3 L-lysine-d-amphetamine 1000 100
[0326] Key observations of this study include: [0327] All animals
in Groups 1-3 showed no observable signs throughout the conduct of
the study. [0328] All animals in Groups 4-6 exhibited increased
motor activity within two hours post-dose and which lasted into Day
2. [0329] One female rat dosed at 1000 mg/kg was found dead on Day
2. Necropsy revealed chromodacryorrhea, chromorhinorrhea, distended
stomach (gas), enlarged adrenal glands, and edematous and distended
intestines. [0330] A total of 4 rats had skin lesions of varying
degrees of severity on Day 3. [0331] One male rat dosed at 1000
mg/kg was euthanatized on Day 3 due to open skin lesions on the
ventral neck. [0332] All remaining animals appeared normal from Day
4 through Day 7.
[0333] Animals were observed for signs of toxicity at 1, 2, and 4 h
post-dose, and once daily for 7 days after dosing and cage-side
observations were recorded. Animals found dead, or sacrificed
moribund were necropsied and discarded.
[0334] Cage-side observations and gross necropsy findings are
summarized above. The oral LD50 of d-amphetamine sulfate is 96.8
mg/kg. For L-lysine-d-amphetamine dimesylate, although the data are
not sufficient to establish a lethal dose, the study indicates that
the lethal oral dose of L-lysine-d-amphetamine is above 1000 mg/kg
because only one death occurred out of a group of six animals.
Although a second animal in this dose group was euthanatized on Day
3, it was done for humane reasons and it was felt that this animal
would have fully recovered. Observations suggested drug-induced
stress in Groups 4-6 that is characteristic of amphetamine toxicity
(NTP, 1990; NIOSH REGISTRY NUMBER: SI1750000; Goodman et. al.,
1985). All animals showed no abnormal signs on Days 4-7 suggesting
full recovery at each treatment level.
[0335] The lack of data to support an established lethal dose is
believed to be due to a putative protective effect of conjugating
amphetamine with lysine. Intact L-lysine-d-amphetamine has been
shown to be inactive, but becomes active upon metabolism into the
unconjugated form (d-amphetamine). Thus, at high doses, saturation
of metabolism of L-lysine-d-amphetamine into the unconjugated form
may explain the lack of observed toxicity, which was expected at
doses greater than 100 mg/kg, which is consistent with
d-amphetamine sulfate (NTP, 1990). The formation rate of
d-amphetamine and the extent of the formation of amphetamine may
both attribute to the reduced toxicity. Alternatively, oral
absorption of L-lysine-d-amphetamine may also be saturated at such
high concentrations, which may suggest low toxicity due to limited
bioavailability of L-lysine-d-amphetamine.
Example 22
In Vitro Assessment of L-lysine-d-amphetamine diHCl Pharmacodynamic
Activity
[0336] It was anticipated that the acylation of amphetamine, as in
the amino acid conjugates discussed here, would significantly
reduce the stimulant activity of the parent drug. For example,
Marvola (1976) showed that N-acetylation of amphetamine completely
abolished the locomotor activity increasing effects in mice. To
confirm that the conjugate was not directly acting as a stimulant,
we tested (NovaScreen, Hanover, Md.) the specific binding of
Lys-Amp (10.sup.-9 to 10.sup.-5 M) to human recombinant dopamine
and norepinephrine transport binding sites using standard
radioligand binding assays. The results (Table 54) indicate that
the Lys-Amp did not bind to these sites. It seems unlikely that the
conjugate retains stimulant activity in light of these results.
(Marvola M. (1976) "Effect of acetylated derivatives of some
sympathomimetic amines on the acute toxicity, locomotor activity
and barbiturate anesthesia time in mice." Acta Pharmacol Toxicol
(Copenh) 38(5): 474-89).
TABLE-US-00059 TABLE 54 Results from radioligand binding
experiments with L-lysine-d-amphetamine Reference Ki (M) for Assay
Radioligand Compound Ref. Cpd. Activity* NE Transporter
[3H]-Nisoxetine Desipramine 4.1 .times. 10.sup.-9 No DA Transporter
[3H]-WIN35428 GBR-12909 7.7 .times. 10.sup.-9 No *No activity is
defined as producing between -20% and 20% inhibition of radioligand
binding (Novascreen).
TABLE-US-00060 TABLE 55 Percent inhibition of DAT and NET with
L-lysine-d-amphetamine L-lysine-d-amphetamine (mol/L) % inhibition
DAT % inhibition NET 10.sup.-9 -10.46 8.15 10.sup.-7 11.52 -11.75
10.sup.-5 -0.71 13.89
Example 23
In Vitro Assessment to Release Amphetamine from
L-lysine-d-amphetamine Dimesylate
[0337] "Kitchen tests" were performed in anticipation of attempts
by illicit chemists to release free amphetamine from the
amphetamine conjugate. Preferred amphetamine conjugates are
resistant to such attempts. Initial kitchen tests assessed the
amphetamine conjugates' resistance to water, acid (vinegar), and
base (baking powder and baking soda) where in each case, the sample
was heated to boiling for 20-60 minutes. L-lysine-d-amphetamine and
GGG-Amp released no detectable free amphetamine.
TABLE-US-00061 TABLE 56 In vitro assessment Vinegar Tap Water
Baking Powder Baking Soda L-lysine-d- 0% 0% 0% 0% amphetamine
Gly.sub.3-Amp 0% 0% 0% 0%
[0338] Amphetamine conjugate stability was assessed under
concentrated conditions, including concentrated HCl and in 10 N
NaOH solution at elevated temperatures. Lys-Amp stock solutions
were prepared in H.sub.2O and diluted 10-fold with concentrated HCl
to a final concentration of 0.4 mg/mL and a final volume of 1.5 mL.
Samples were heated in a water bath to about 90.degree. C. for 1
hour, cooled to 20.degree. C., neutralized, and analyzed by HPLC
for free d-amphetamine. The results suggest that only a minimal
amount of d-amphetamine is released under these concentrated
conditions.
TABLE-US-00062 TABLE 57 Stability under concentrated conditions %
AUC solution Lys-Amp d-amphetamine 10 N NaOH 99 <1 conc. HCl 96
4
[0339] Amphetamine conjugate stability was assessed under acidic
conditions.
TABLE-US-00063 TABLE 58 Acids used for stability study Acid
Concentrations Hydrochloric acid 10%, 25%, 50%, 75%, and
concentrated Acetic acid 10%, 25%, 50%, 75%, and concentrated
Sulfuric acid 10%, 25%, 50%, 75%, and concentrated Phosphoric acid
10%, 25%, 50%, 75%, and concentrated Nitric acid 10%, 25%, 50%,
75%, and concentrated Citric acid 10%, 25%, 50%, 75%, and
saturated
[0340] At ambient temperature, only a limited amount of
d-amphetamine was released. At 90.degree. C., only a limited amount
of d-amphetamine was released, but the decomposition of
L-lysine-d-amphetamine was more pronounced. This suggested that the
amide bond is stable, and that the conjugate usually degrades
before an appreciable amount is hydrolyzed. At reflux conditions,
concentrated hydrochloric acid and 50% sulfuric acid released 85%
and 59%, respectively, of the d-amphetamine content, but rendered
the drug in undesirable acidic solution. The process for recovering
d-amphetamine from the acidic solution further reduces the
yield.
[0341] In a similar test, reflux in concentrated HCl resulted in
some hydrolysis after 5 hours (28%) with further hydrolysis
occurring after 22 hours (76%). Reflux in concentrated
H.sub.2SO.sub.4 for 2 hours resulted in complete decomposition of
Lys-Amp and potentially released d-amphetamine. As described above,
recovery of d-amphetamine from the acidic solution would further
reduce the yield.
[0342] Amphetamine conjugate stability was also assessed under
basic conditions, including variable concentrations of sodium
hydroxide, potassium hydroxide, sodium carbonate, ammonium
hydroxide, diethyl amine, and triethyl amine. The maximum
d-amphetamine release was 25.4% obtained by 3M sodium hydroxide;
all other basic conditions resulted in a release of less than
3%.
Example 24
Stability of L-lysine-d-amphetamine Dimesylate Under Treatment with
Commercially Available Products
[0343] The stability of L-lysine-d-amphetamine dimesylate was
assessed under treatment commercially available acids, bases, and
enzyme cocktails. For acids and bases (Table 59), 10 mg of Lys-Amp
was mixed with 2 mL of each stock solution, and the solution was
shaken at 20.degree. C. For enzyme treatment (Table 60), 10 mg
Lys-Amp was mixed with 5 mL of each enzyme cocktail, and the
solution was shaken at 37.degree. C. Each aliquot (0, 1, and 24 h)
was neutralized and filtered prior to analysis by HPLC. Many of the
commercially available reagents also contained various solvents
and/or surfactants.
[0344] Unless otherwise indicated, solutions were used directly
from the container and were combined with neat Lys-Amp solid. Lewis
Red Devil.RTM. Lye, Enforcer Drain Care.RTM. Septic Treatment, and
Rid-X.RTM. Septic Treatment were prepared as saturated solutions in
H.sub.2O. Enzymes used were purchased from Sigma and directly
dissolved in water (3 mg/mL pepsin, 10 mg/mL pancreatin, 3 mg/mL
pronase, 3 mg/mL esterase), while enzyme-containing nutraceuticals
such as Omnigest.RTM. and VitalZym.RTM. were first either crushed
or opened (1 tablet or capsule per 5 mL of H.sub.2O).
[0345] The commercial acids and bases were ineffective in
hydrolyzing Lys-Amp. Only treatment with Miracle-Gro.RTM. (7%
release) and Olympic.RTM. Deck Cleaner (4% release) showed any
release, but even after 24 hours, the amount of d-amphetamine was
negligible. Among the enzyme products, only pure esterase (19%
release) or pronase (24% release) mixtures successfully cleaved
lysine (after 24 hours).
TABLE-US-00064 TABLE 59 Stability of L-lysine-d-amphetamine
dimesylate under treatment with commercially available acids and
bases d-amphetamine (1) (w.t. % d-amphetamine) Solution (active
ingredients) 1 h 24 h Lysol .RTM. Toilet (HCl) 0 0 Crete-nu (75%
H.sub.3PO.sub.4) 0 0 CLR .RTM. (sulfamic acid, hydroxyacetic acid)
0 0 Roebic .RTM. Drain Flow (90% H.sub.2SO.sub.4) 0 0 Crown .RTM.
Muriatic Acid (31.45% HCl) 0 0 Liquid-Plumr .RTM. (NaOH, NaClO,
H.sub.2O.sub.2) 0 0 Brasso .RTM. (NH.sub.4OH) 0 0 Johnson .RTM. Wax
Degreaser (K.sub.2CO.sub.3) 0 0 Miracle-Gro .RTM. (Urea,
K.sub.3PO.sub.4) 0 7 Lewis Red Devil .RTM. Lye (NaOH) 0 0 Drain
Power (NaOH, NaClO) 0 0 Savogran TSP (Na.sub.3PO.sub.4) 0 0 Johnson
.RTM. Wax Stripper (NaOH) 0 0 Olympic .RTM. Deck Cleaner (NaOH,
NaClO) 0 4 Windex .RTM. (NH.sub.4OH) 0 0 Greased Lightning .RTM.
(basic components) 0 0
TABLE-US-00065 TABLE 60 Stability of L-lysine-d-amphetamine
dimesylate under treatment with commercially available enzyme
cocktails d-amphetamine (1) (w.t. % d-amphetamine) Solution 1 h 24
h Cellfood .RTM. 0 0 Drano .RTM. Max with Bacteria 0 0 VitalZym
.RTM. 0 0 Omnigest .RTM. 0 0 Enforcer .RTM. Septic 0 0 Rid-X .RTM.
Septic 0 0 Esterase 0 19 Pancreatin 0 0 Pepsin 0 0 Pronase 0 24
Example 25
Bioavailability of Various Peptide Amphetamine Conjugates (HCl
Salts) Administered by Oral, Intranasal, and Intravenous Routes
[0346] Oral administration: Male Sprague-Dawley rats were provided
water ad libitum, fasted overnight, and dosed by oral gavage with
amphetamine or amino acid-amphetamine conjugates containing the
equivalent amount of amphetamine.
[0347] Intranasal administration: Male Sprague-Dawley rats were
dosed by intranasal administration with amphetamine or
lysine-amphetamine (1.8 mg/kg).
[0348] The relative in vivo performance of various amino
acid-amphetamine compounds is shown in FIG. 50-FIG. 58 and
summarized in Table 61. Intranasal bioavailability of amphetamine
from Ser-Amp was decreased to some degree relative to free
amphetamine. However, this compound was not bioequivalent with
amphetamine by the oral route of administration. Phenylalanine was
bioequivalent with amphetamine by the oral route of administration,
however, little or no decrease in bioavailability by parenteral
routes of administration was observed. Gly.sub.3-Amp had nearly
equal bioavailability (90%) by the oral route accompanied by a
decrease in C.sub.max (74%). Additionally, Gly.sub.3-Amp showed a
decrease in bioavailability relative to amphetamine by intranasal
and intravenous routes.
TABLE-US-00066 TABLE 61 Percent bioavailability of amino acid
amphetamine compounds administered by oral, intranasal, or
intravenous routes Oral Intranasal Intravenous Percent Percent
Percent Percent Percent Percent Drug AUC C.sub.max AUC C.sub.max
AUC C.sub.max Amphetamine 100 100 100 100 100 100 E-Amp 73 95 NA NA
NA NA EE-Amp 26 74 NA NA NA NA EEE-Amp 69 53 10 10 NA NA L-Amp 65
81 NA NA NA NA S-Amp 79/55 62/75 76 65 NA NA G-Amp 81 78 65 53 NA
NA GG-Amp 79 88 88 85 NA NA GGG-Amp 111/68 74/73 32 38 45 46 F-Amp
95 91 97 95 87 89 EEF-Amp 42 73 39 29 NA NA FF-Amp 27 64 NA NA NA
NA Gulonate-Amp 1 1 0.4 0.5 3 5 K-Amp 98 55 0.5 0.5 3 3 KG-Amp 69
71 13 12 NA NA dKlK-Amp 16 7 2 2 NA NA LE-Amp 40 28 6 6 NA NA H-Amp
16 21 22 42 NA NA P-Amp 6 3 2 2 NA NA PP-Amp 61 80 47 43 NA NA
Y-Amp 25 20 21 20 NA NA I-Amp 71 52 73 97 NA NA
[0349] Several single amino acid amphetamine conjugates had
comparable oral bioavailability (80-100%) to d-amphetamine. Lys,
Gly, and Phe conjugates, for example, all demonstrated similar oral
bioavailability to the parent drug. Dipeptide prodrugs generally
showed lower bioavailability than the respective amino acid
analogs, and tripeptide compounds displayed no discernable trend.
Several amino acid amphetamine conjugates had decreased parenteral
bioavailability. Preferred conjugates, such as Lys-Amp, exhibit
both oral bioavailability comparable to d-amphetamine and decreased
parenteral bioavailability compared to d-amphetamine.
Example 26
Decreased Oral C.sub.max of d-amphetamine Conjugates
[0350] Male Sprague-Dawley rats were provided water ad libitum,
fasted overnight, and dosed by oral gavage with amphetamine
conjugate or d-amphetamine sulfate. All doses contained equivalent
amounts of d-amphetamine base. Plasma d-amphetamine concentrations
were measured by ELISA (Amphetamine Ultra, 109319, Neogen,
Corporation, Lexington, Ky.). The assay is specific for
d-amphetamine with only minimal reactivity (0.6%) of the major
d-amphetamine metabolite (para-hydroxy-d-amphetamine) occurring.
Plasma d-amphetamine and L-lysine-d-amphetamine concentrations were
measured by LC/MS/MS where indicated in examples.
Example 27
Decreased Intranasal Bioavailability (AUC and C.sub.max) of
d-amphetamine Conjugates
[0351] Male Sprague-Dawley rats were provided water ad libitum and
doses were administered by placing 0.02 ml of water containing
amphetamine conjugate or d-amphetamine sulfate into the nasal
flares. All doses contained equivalent amounts of d-amphetamine
base. Plasma d-amphetamine concentrations were measured by ELISA
(Amphetamine Ultra, 109319, Neogen, Corporation, Lexington, Ky.).
The assay is specific for d-amphetamine with only minimal
reactivity (0.6%) of the major d-amphetamine metabolite
(para-hydroxy-d-amphetamine) occurring. Plasma d-amphetamine and
L-lysine-d-amphetamine concentrations were measured by LC/MS/MS
where indicated in examples.
Example 28
Decreased Intravenous Bioavailability (AUC and C.sub.max) of
d-amphetamine Conjugates
[0352] Male Sprague-Dawley rats were provided water ad libitum, and
doses were administered by intravenous tail vein injection of 0.1
ml of water containing amphetamine conjugate or d-amphetamine
sulfate. All doses contained equivalent amounts of d-amphetamine
base. Plasma d-amphetamine concentrations were measured by ELISA
(Amphetamine Ultra, 109319, Neogen, Corporation, Lexington, Ky.).
The assay is specific for d-amphetamine with only minimal
reactivity (0.6%) of the major d-amphetamine metabolite
(para-hydroxy-d-amphetamine) occurring. Plasma d-amphetamine and
L-lysine-d-amphetamine concentrations were measured by LC/MS/MS
where indicated in examples.
Example 29
Attachment of Amphetamine to Variety of Chemical Moieties
[0353] The above examples demonstrate the use of an amphetamine
conjugated to a chemical moiety, such as an amino acid, which is
useful in reducing the potential for overdose while maintaining its
therapeutic value. The effectiveness of binding amphetamine to a
chemical moiety was demonstrated through the attachment of
amphetamine to lysine (K), however, the above examples are meant to
be illustrative only. The attachment of amphetamine to any variety
of chemical moieties (i.e., peptides, glycopeptides, carbohydrates,
nucleosides, or vitamins) as described below through similar
procedures using the following exemplary starting materials.
Amphetamine Synthetic Examples
Synthesis of Gly.sub.2-Amp
[0354] Gly.sub.2-Amp was synthesized by a similar method except the
amino acid starting material was Boc-Gly-Gly-OSu.
[0355] Synthesis of Glu.sub.2-Phe-Amp [0356] Glu2-Phe-Amp was
synthesized by a similar method except the amino acid starting
material was Boc-Glu(OtBu)-Glu(OtBu)-OSu and the starting drug
conjugate was Phe-Amp (see Phe-Amp synthesis).
[0357] Synthesis of His-Amp
[0358] His-Amp was synthesized by a similar method except the amino
acid starting material was Boc-His(Trt)-OSu.
[0359] Synthesis of Lys-Gly-Amp
[0360] Lys-Gly-Amp was synthesized by a similar method except the
amino acid starting material was Boc-Lys(Boc)-OSu and the starting
drug conjugate was Gly-Amp (see Gly-Amp synthesis).
[0361] Synthesis of Lys-Glu-Amp
[0362] Lys-Glu-Amp was synthesized by a similar method except the
amino acid starting material was Boc-Lys(Boc)-OSu and the starting
drug conjugate was Glu-Amp.
[0363] Synthesis of Glu-Amp
[0364] Glu-Amp was synthesized by a similar method except the amino
acid starting material was Boc-Glu(OtBu)-OSu.
[0365] Synthesis of (d)-Lys-(l)-Lys-Amp
[0366] (d)-Lys-(l)-Lys-Amp was synthesized by a similar method
except the amino acid starting material was
Boc-(d)-Lys(Boc)-(l)-Lys(Boc)-OSu.
[0367] Synthesis of Gulonic acid-Amp
[0368] Gul-Amp was synthesized by a similar method except the
carbohydrate starting material was gulonic acid-OSu.
Example 30
Lack of Detection of L-lysine-d-amphetamine diHCl in Brain Tissue
Following Oral Administration
[0369] Male Sprague-Dawley rats were provided water ad libitum,
fasted overnight, and dosed by oral gavage with
L-lysine-d-amphetamine or d-amphetamine sulfate. All doses
contained equivalent amounts of d-amphetamine base. As shown in
FIG. 59, similar levels of d-amphetamine were detected in serum as
well as in brain tissue following administration of d-amphetamine
sulfate or L-lysine-d-amphetamine. The d-amphetamine from
L-lysine-d-amphetamine showed a sustained presence in the brain as
compared to levels of d-amphetamine from d-amphetamine sulfate. The
conjugate L-lysine-d-amphetamine was present in appreciable amounts
in serum but was not detected in brain tissue indicating that the
conjugate does not cross the blood brain barrier to access the
central nervous system site of action.
Example 31
Pharmaceutical Composition of L-lysine-d-amphetamine Dimesylate
[0370] A gelatin capsule dosage form was prepared in three dosage
strengths. The hard gelatin capsules were printed with NRP104 and
the dosage strength. The capsule fill contains a white to off-white
finely divided powder uniform in appearance.
TABLE-US-00067 TABLE 62 Composition of L-lysine-d-amphetamine
dimesylate capsules Quantity (mg) Ingredient 30 50 70 Placebo
Function Grade L-lysine-d- 30.0 50.0 70.0 0.0 Active amphetamine
dimesylate Microcrystalline 151 70.0 98.0 144.0 Filler/ NF (Avicel
.RTM. Cellulose diluent, PH-102) disintegrant Croscarmellose 4.69
3.12 4.37 3.75 Disintegrant NF Sodium Magnesium 1.88 1.88 2.63 2.25
Lubricant NF (5712) Stearate Gelatin Capsule White/ White/ Med.
White/ Carrier NF Size 3 Med. Lt. Orange/ White Orange Blue Lt.
Blue Total 187.5 125 175 150
[0371] Other diluents, disintegrants, lubricants, and colorants,
etc. may be used. Also, a particular ingredient can be used to
serve a different function than those listed above.
[0372] The pharmaceutical composition was prepared by milling
de-lumped L-lysine-d-amphetamine dimesylate (size 20 mesh) with
microcrystalline cellulose. The mixture was sieved through a 30
mesh screen and then mixed with croscarmellose sodium. Pre-screened
magnesium stearate (size 30 mesh) was added, and the composition
was mixed until uniform to form the capsule fill.
Example 32
Clinical Pharmacokinetic Evaluation and Oral Bioavailability of
L-lysine-d-amphetamine Dimesylate 70 mg Capsules Administered to
Healthy Adults Under Fasting Conditions for 7 Days
[0373] In this open-label, single-arm study, healthy adults between
the ages of 18 to 55 years were administered 70 mg of
L-lysine-d-amphetamine dimesylate with 8 ounces of water once daily
(7 am) for 7 consecutive days. Patients fasted for at least 10
hours before and 4 hours after final dosing. Venous blood samples
(7 mL) were drawn into EDTA vacutainers both before medication
dosing on days 0, 1, 6, and 7 (in the morning) and at 16 time
points (hours 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 10, 12, 16, 24, 48,
and 72) after final dosing on day 7. Immediately after sample
collection, vacutainer tubes were centrifuged at 3000 rpm at
4.degree. C. for 10 minutes; within 1 hour of collection, they were
stored at -20.degree. C. Plasma samples were analyzed for
L-lysine-d-amphetamine and d-amphetamine using a validated LC/MS/MS
method.
[0374] By dose 5, d-amphetamine reached steady state. After dose 7,
mean AUC.sub.0-24 was 1113 ngh/mL, mean AUC.sub.0-.infin. was 1453
ngh/mL, mean C.sub.max was 90.1 ngh/mL, and mean T.sub.max was 3.68
hours. See Table 63 and FIG. 60. In comparison, extended-release
amphetamine salts exhibit a T.sub.max of 5.8 hours and
AUC.sub.0-.infin. 853 ngh/mL after an overnight fast. J. F. Auiler
et al., "Effect of food on early drug exposure from
extended-release stimulants: results from the Concerta, Adderall XR
Food Evaluation (CAFE) study," Curr Med Res Opin 18: 311-316 at 313
(2002).
[0375] Intact L-lysine-d-amphetamine was rapidly converted to
d-amphetamine. After dose 7, mean AUC.sub.0-24 was 60.66 ngh/mL,
and mean AUC.sub.0-.infin. was 61.06 ngh/mL. See Table 63 and FIG.
60. In addition, mean C.sub.max was 47.9 ngh/mL, and mean T.sub.max
was 1.14 hours for intact L-lysine-d-amphetamine.
L-lysine-d-amphetamine was completely eliminated within
approximately 6 hours.
[0376] The results also indicate that l-lysine-d-amphetamine
administration leads to a decrease in patient to patient
variability of amphetamine levels (see the lower CV values for the
L-lysine-d-amphetamine conjugate as compared to d-amphetamine).
[0377] There were no gender differences in systemic exposure to
d-amphetamine, though C.sub.max was 12% higher in men after
normalization by body weight.
[0378] The multidose pharmacokinetic profile of d-amphetamine
released from the prodrug L-lysine-d-amphetamine is consistent with
extended-release properties. The adverse events that occurred in
this setting are consistent with other stimulants and suggest that
suggest that L-lysine-d-amphetamine 70 mg is well tolerated.
TABLE-US-00068 TABLE 63 Steady-state pharmacokinetics parameters (n
= 11) Parameter Mean SD CV % d-amphetamine C.sub.max (ng/mL) 90.1
29.6 32.84 C.sub.min (ng/mL) 18.2 14.2 78.12 T.sub.max (h) 3.68
1.42 38.54 t.sub.1/2 (h) 10.08 2.76 27.37 AUC.sub.0-24 (ng h/mL)
1113 396.8 35.65 AUC.sub.0-.infin. (ng h/mL) 1453 645.7 44.45
AUC.sub.0-t (ng h/mL) 1371 633.5 46.19 FI (%) 163.55 37.20 22.74
Intact L-lysine-d-amphetamine C.sub.max (ng/mL) 47.9 18.6 38.81
C.sub.min (ng/mL) 0.0 0.0 -- T.sub.max (h) 1.14 0.32 28.45
t.sub.1/2 (h) 0.43 0.09 21.90 AUC.sub.0-24 (ng h/mL) 60.66 21.00
34.61 AUC.sub.0-.infin. (ng h/mL) 61.06 20.63 33.79 AUC.sub.0-t (ng
h/mL) 59.44 21.47 36.12 FI (%) 1896.06 340.24 17.94
Example 33
Clinical Pharmacokinetic Evaluation and Oral Bioavailability of
L-lysine-d-amphetamine Dimesylate Compared to Amphetamine Extended
Release Products Adderall XR.RTM. and Dexedrine Spansule.RTM. Used
in the Treatment of ADHD
TABLE-US-00069 [0379] TABLE 64 Treatment groups and dosage for
clinical pharmacokinetic evaluation of L- lysine-d-amphetamine
compared to Adderall XR .RTM. or Dexedrine Spansule .RTM. Treatment
No. of Dose Dose Drug Group Subjects Dose (mg) (amphetamine base)
L-lysine- A 10 1 .times. 25 mg 25 7.37 d-amphetamine capsule
L-lysine- B 10 3 .times. 25 mg capsules 75 22.1 d-amphetamine
Dexedrine C 10 3 .times. 10 mg capsules 30 22.1 Spansule .RTM.
Adderall XR .RTM. D 10 1 .times. 30 mg capsules 35 21.9 plus 1
.times. 5 mg capsule
[0380] A clinical evaluation of the pharmacokinetics and oral
bioavailability of L-lysine-d-amphetamine in humans was conducted.
L-lysine-d-amphetamine was orally administered at doses
approximating the lower (25 mg) and higher (75 mg) end of the
therapeutic range based on d-amphetamine base content of the doses.
Additionally, the higher dose was compared to doses of Adderall
XR.RTM. (Shire) or Dexedrine Spansule.RTM. (GlaxoSmithKline)
containing equivalent amphetamine base to that of the higher
L-lysine-d-amphetamine dose. Treatment groups and doses are
summarized in Table 64. All levels below limit quantifiable (blq
<0.5 ng/mL) were treated as zero for purposes of pharmacokinetic
analysis.
[0381] The concentrations of d-amphetamine and
L-lysine-d-amphetamine intact conjugate following administration of
L-lysine-d-amphetamine at the low and high dose for each individual
subject as well as pharmacokinetic parameters are presented in
Table 65-Table 70. The concentrations of d-amphetamine following
administration of Adderall XR.RTM. or Dexedrine Spansule.RTM. for
each individual subject as well as pharmacokinetic parameters are
presented in Table 69 and Table 70, respectively.
Concentration-time curves showing L-lysine-d-amphetamine intact
conjugate and d-amphetamine are presented in FIG. 61 and FIG. 62.
Extended release of d-amphetamine from L-lysine-d-amphetamine was
observed for both doses and pharmacokinetic parameters (C.sub.max
and AUC) were proportional to doses when the lower and higher dose
results were compared (FIG. 61 and FIG. 62). Significant levels of
d-amphetamine were not observed until one-hour post administration.
Only small amounts (1.6 and 2.0 percent of total drug absorption,
respectively for 25 and 75 mg doses; AUC.sub.inf-molar basis) of
L-lysine-d-amphetamine intact conjugate were detected with levels
peaking at about one hour (Table 66 and Table 68). The small amount
of intact conjugate absorbed was rapidly and completely eliminated,
with no detectable concentrations present by five hours, even at
the highest dose.
[0382] In a cross-over design (identical subjects received Adderall
XR.RTM. doses following a 7-day washout period), the higher
L-lysine-d-amphetamine dose was compared to an equivalent dose of
Adderall XR.RTM.. Adderall XR.RTM. is a once-daily extended release
treatment for ADHD that contains a mixture of d-amphetamine and
l-amphetamine salts (equal amounts of d-amphetamine sulfate,
d-/l-amphetamine sulfate, d-amphetamine saccharate, and
d-/l-amphetamine aspartate). An equivalent dose of extended release
Dexedrine Spansule.RTM. (contains extended release formulation of
d-amphetamine sulfate) was also included in the study. As observed
in pharmacokinetic studies in rats, oral administration of
L-lysine-d-amphetamine resulted in d-amphetamine concentration-time
curves similar to those of Adderall XR.RTM. and Dexedrine
Spansule.RTM. (FIG. 63 and FIG. 64). The bioavailability
(AUC.sub.inf) of d-amphetamine following administration of
L-lysine-d-amphetamine was approximately equivalent to both
extended release amphetamine products (Table 71). Over the course
of twelve hours, typically the time needed for effective once-daily
treatment of ADHD, the bioavailability for L-lysine-d-amphetamine
was approximately equivalent to that of Adderall XR.RTM.
(d-amphetamine plus l-amphetamine levels) and over twenty percent
higher than that of Dexedrine Spansule.RTM.. Based on the results
of this clinical study, L-lysine-d-amphetamine would be an
effective once-daily treatment for ADHD. Moreover,
L-lysine-d-amphetamine afforded similar pharmacokinetics in humans
and animal models, namely, delayed release of d-amphetamine
resulting in extended release kinetics. Based on these observations
L-lysine-d-amphetamine should also have abuse-resistant properties
in humans.
TABLE-US-00070 TABLE 65 Individual subject d-amphetamine
concentrations and pharmacokinetic parameters following oral
administration of a 25 mg dose of L-lysine-d-amphetamine to humans
Subject Subject Subject Subject Subject Subject Subject Subject
Subject Subject 102 103 105 107 110 112 113 116 117 120 Mean SD CV
% Time Hours 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.5 0 0 0.625 0 0 0 0 0.78
0.769 0 0.2 0.4 162.1 1 4.29 2.95 8.67 3.36 8.33 1.1 10 10.5 14
3.15 6.6 4.2 63.6 1.5 10 12.7 16 13.8 21.4 3.94 24.7 19.5 24 15.1
16.1 6.5 40.3 2 16.3 18.4 17 21 25.9 9.29 30.9 23.6 30 21.7 21.4
6.6 30.8 3 16.5 19.6 16.7 26.1 27 17.7 30.2 23.5 27.6 28.9 23.4 5.3
22.7 4 23.9 18.8 14.1 24.5 30.1 17.9 33.2 21.2 24.7 25.3 23.4 5.7
24.3 5 21.2 18.9 14.6 21.6 22.6 17.2 27 20 20.2 24.2 20.8 3.5 16.9
6 21.8 18 12.5 21.6 23.7 15.7 25.8 18.2 20.3 20.5 19.8 3.9 19.6 7
18.9 15.8 12.1 17.8 20.6 14.5 26.6 21 18.3 21.8 18.7 4.1 21.9 8
19.3 16.6 10.4 17.9 20 14.2 25.7 13.6 18.8 20.1 17.7 4.2 24.1 10
18.8 13.6 9.8 15.3 19.3 13.7 22.4 15.1 15.3 15.9 15.9 3.5 22.1 12
15.8 12.6 6.92 11.5 15.8 11.2 17.9 12 13.7 15.2 13.3 3.1 23.6 16
13.4 10.5 6.56 9.53 14.3 10.7 12.5 10.3 10 13 11.1 2.3 20.5 24 8.03
5.81 2.65 4.9 5.8 5.9 6.57 6.13 4.52 5.45 5.6 1.4 25.1 48 1.57 1.36
0 1.26 0.795 1.44 1.24 1.23 0.864 0.586 1.0 0.5 46.1 72 0 0 0 0 0 0
0 0 0 0 0 0 0 Parameter AUC.sub.0-12 h 204.0 177.4 140.4 204.9
242.7 152.4 284.6 199.2 225.5 223.3 205.4 42.5 20.7 (ng h/mL)
AUC.sub.last 463.3 375.1 201.4 378.5 462.7 350.7 515.2 397.9 395.7
426.1 396.7 84.8 21.4 (ng h/mL) AUC.sub.inf 486.7 397.1 233.5 398.8
472 374 532.5 416.4 407 432.2 415.0 80.1 19.3 (ng h/mL) C.sub.max
23.9 19.6 17 26.1 30.1 17.9 33.2 23.6 30 28.9 25.0 5.6 22.3 (ng/mL)
T.sub.max (hours) 4 3 2 3 4 4 4 2 2 3 3.1 0.876 28.2 T.sub.1/2
(hours) 10.32 11.18 8.36 11.18 8.16 11.22 9.68 10.43 9.06 7.22 9.68
1.43 14.7
TABLE-US-00071 TABLE 66 Individual subject L-lysine-d-amphetamine
intact conjugate concentrations and pharmacokinetic parameters
following oral administration of a 25 mg dose of
L-lysine-d-amphetamine to humans Subject Subject Subject Subject
Subject Subject Subject Subject Subject Subject 102 103 105 107 110
112 113 116 117 120 Mean SD CV % Time Hours 0 0 0 0 0 0 0 0 0 0 0 0
0 0 0.5 4.1 5.5 10.0 0.0 3.6 0.0 9.2 9.6 8.9 0.0 5.1 4.2 82.0 1 9.2
11.2 15.2 12.5 9.1 2.7 20.1 10.5 10.8 10.9 11.2 4.5 39.7 1.5 4.0
4.4 6.1 7.5 3.6 6.2 6.6 2.8 4.2 8.4 5.4 1.8 34.1 2 2.1 1.4 2.5 2.9
1.9 4.0 2.3 0 1.7 3.1 2.2 1.1 48.8 3 0 0 0 0 0 0 0 0 0 0 0 0 0 4 0
0 0 0 0 0 0 0 0 0 0 0 0 5 0 0 0 0 0 0 0 0 0 0 0 0 0 6 0 0 0 0 0 0 0
0 0 0 0 0 0 7 0 0 0 0 0 0 0 0 0 0 0 0 0 8 0 0 0 0 0 0 0 0 0 0 0 0 0
10 0 0 0 0 0 0 0 0 0 0 0 0 0 12 0 0 0 0 0 0 0 0 0 0 0 0 0 16 0 0 0
0 0 0 0 0 0 0 0 0 0 24 0 0 0 0 0 0 0 0 0 0 0 0 0 48 0 0 0 0 0 0 0 0
0 0 0 0 0 72 0 0 0 0 0 0 0 0 0 0 0 0 0 Parameter AUC.sub.last 9.18
10.95 16.31 10.68 8.583 5.439 18.51 10.77 12.35 10.41 11.32 3.74
33.1 (ng h/mL) AUC.sub.inf 10.62 11.64 17.66 12.65 9.759 -- 19.56
-- 13.3 12.83 13.50 3.40 25.2 (ng h/mL) C.sub.max (ng/mL) 9.18 11.2
15.2 12.5 9.05 6.18 20.1 10.5 10.8 10.9 11.56 3.80 32.9 T.sub.max
(hours) 1 1 1 1 1 1.5 1 1 1 1 1.05 0.16 15.1 T.sub.1/2 (hours) 0.47
0.34 0.38 0.47 0.44 -- 0.32 -- 0.38 0.55 0.419 0.077 18.5
TABLE-US-00072 TABLE 67 Individual subject d-amphetamine
concentrations and pharmacokinetic parameters following oral
administration of a 75 mg dose of L-lysine-d-amphetamine to humans
Subject Subject Subject Subject Subject Subject Subject Subject
Subject Subject 101 104 106 108 109 111 114 115 118 119 Mean SD CV
% Time Hours 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.5 0 0.748 0.506 0 0
0.779 0.525 0 3 1.85 0.7 1.0 132.2 1 11.9 14.4 12.6 7.26 5.9 10.3
7.2 23.1 23 27.9 14.4 7.7 53.6 1.5 40.3 34.6 30.4 22.8 19.3 38.4 19
52.8 51.5 55.8 36.5 13.8 37.8 2 84.6 48.9 68.2 34.8 32.7 57.2 33.1
91.3 61.7 70.4 58.3 21.0 36.0 3 72.9 64.3 55.7 60.3 62.3 61.1 44.8
95.8 62.1 83.6 66.3 14.5 21.9 4 84.6 65.3 58.8 51.1 77.9 63.3 47.6
89.2 54.2 86 67.8 15.5 22.8 5 65 55.6 60.2 74 83.9 59.1 56.9 77.7
54.9 82.8 67.0 11.5 17.2 6 71 53.5 49.4 51.5 78.3 50.8 55.1 68.8
52.9 64 59.5 10.2 17.1 7 53.8 55.7 52.9 69.5 73.1 52.9 55.9 71.2
45.1 74.6 60.5 10.5 17.4 8 63.7 40.3 47.3 45.7 72.2 46.5 54.2 61.1
44.3 66.2 54.2 10.9 20.2 10 43.7 41.7 37 58.4 67 44.3 48.4 68 34.1
55.9 49.9 11.9 24.0 12 46.4 26.1 36.7 37.4 49.9 32.4 37.1 54.1 34.5
45.1 40.0 8.6 21.6 16 35.4 22.2 25.7 48 44.9 24.3 28.9 44.7 31.7
34.5 34.0 9.2 27.1 24 16.4 11.4 14.9 13.2 18.4 16.8 20.5 21.7 15.7
18.1 16.7 3.1 18.8 48 2.74 2.14 4.17 2.73 3.75 4.81 2.81 4.26 3.36
3.4 0.9 25.9 72 0 0 0 1.07 0.661 0.687 1.49 0 0 0.553 0.4 0.5 120.2
Parameter AUC.sub.0-12 h 666.2 525.9 531.6 570.3 704.8 545.6 513.7
790.9 523.4 742.8 611.5 104.5 17.1 (ng h/mL) AUC.sub.last 1266
918.7 1031 1257 1442 1123 1223 1549 1143 1417 1237.0 194.0 15.7 (ng
h/mL) AUC.sub.inf 1301 948.3 1072 1278 1451 1133 1251 1582 1154
1425 1259.5 191.3 15.2 (ng h/mL) C.sub.max (ng/mL) 84.6 65.3 68.2
74 83.9 63.3 56.9 95.8 62.1 86 74.0 12.9 17.4 T.sub.max (hours) 4 4
2 5 5 4 5 3 3 4 3.9 1.0 25.5 T.sub.1/2 (hours) 8.78 9.59 10.02
13.26 9.24 10.41 12.8 8.05 10.92 9.47 10.3 1.7 16.3
TABLE-US-00073 TABLE 68 Individual subject L-lysine-d-amphetamine
intact conjugate concentrations and pharmacokinetic parameters
following oral administration of a 75 mg dose of
L-lysine-d-amphetamine to humans Subject Subject Subject Subject
Subject Subject Subject Subject Subject Subject 101 104 106 108 109
111 114 115 118 119 Mean SD CV % Time Hours 0 0 0 0 0 0 0 0 0 0 0 0
0 0 0.5 10.4 22.6 6.92 10.3 0 9.21 7.88 14.5 87.8 35.5 20.5 25.6
124.7 1 48 40.5 29 41.5 21.2 30.8 23.4 127 88.9 80.1 53.0 34.6 65.2
1.5 28.4 15.7 16.1 20.3 26.5 19 12.7 38.7 28.6 38 24.4 9.2 37.5 2
8.87 5.53 4.91 9 18.1 5.62 6.29 12.1 9.75 11.3 9.1 4.0 44.0 3 2.15
1.29 1.76 1.82 10.6 0 2.31 2.57 1.73 1.73 2.6 2.9 111.6 4 0 0 1.09
0 4.65 0 1.53 1.01 0 0 0.8 1.5 176.9 5 0 0 0 0 0 0 0 0 0 0 0 0 0 6
0 0 0 0 0 0 0 0 0 0 0 0 0 7 0 0 0 0 0 0 0 0 0 0 0 0 0 8 0 0 0 0 0 0
0 0 0 0 0 0 0 10 0 0 0 0 0 0 0 0 0 0 0 0 0 12 0 0 0 0 0 0 0 0 0 0 0
0 0 16 0 0 0 0 0 0 0 0 0 0 0 0 0 24 0 0 0 0 0 0 0 0 0 0 0 0 0 48 0
0 0 0 0 0 0 0 0 0 0 0 0 72 0 0 0 0 0 0 0 0 0 0 0 0 0 Parameter
AUC.sub.last 51.2 44.2 32.0 43.7 50.4 30.9 29.8 102.1 110.8 86.1
58.1 30.2 52.0 (ng h/mL) AUC.sub.inf 52.5 45.0 33.0 44.9 52.3 34.2
31.4 102.9 111.7 87.0 59.5 29.9 50.2 (ng h/mL) C.sub.max 48.0 40.5
29.0 41.5 26.5 30.8 23.4 127.0 88.9 80.1 53.6 34.1 63.6 (ng/mL)
T.sub.max (hours) 1 1 1 1 1.5 1 1 1 1 1 1.05 0.16 15.1 T.sub.1/2
(hours) 0.43 0.4 0.61 0.43 1.02 0.41 0.75 0.56 0.38 0.35 0.534
0.211 39.6
TABLE-US-00074 TABLE 69 Individual subject d-amphetamine
concentrations and pharmacokinetic parameters following oral
administration of a 35 mg dose of Adderall XR .RTM. (equivalent to
75 mg dose of L-lysine-d-amphetamine based on amphetamine base
content) to humans Subject Subject Subject Subject Subject Subject
Subject Subject Subject Subject 101 104 106 108 109 111 114 115 118
119 Mean SD CV % Time Hours 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.5 7.9 2.3
2.8 0.6 2.2 5.7 0 16 2.3 5.3 4.5 4.7 104.3 1 37.6 28.9 23.3 13.7
29.8 38.2 17.9 46.2 28.8 48.8 31.3 11.5 36.6 1.5 49.9 42.3 31.1
23.7 39.1 34.4 30.8 65.4 34.1 53 40.4 12.5 31.0 2 65.9 45.8 29.2
37.4 46.2 65.4 40 64.4 37 67.8 49.9 14.6 29.2 3 95.3 51.7 36.7 23.6
64.7 62.9 44.7 56.5 31.1 64.8 53.2 20.7 38.9 4 83.7 73.3 56.7 40 67
76.6 56.3 53.1 33.5 73.3 61.4 16.3 26.6 5 77.4 75.2 71.6 62.1 75.9
76.4 51.5 61.4 56.8 82.4 69.1 10.3 14.9 6 71.5 72.1 64 59.8 66.9
63.5 56.8 59.8 58.7 85.7 65.9 8.7 13.2 7 72.3 63.6 71 57.9 70.6
69.7 51.9 48.1 53.7 79.7 63.9 10.5 16.4 8 60.4 57.1 53.8 53 72 66.9
56.2 56.4 51.7 66.7 59.4 6.9 11.6 10 50.4 45.5 53 50.7 67.6 57.4
49.1 66.6 48 71.3 56.0 9.3 16.6 12 42.5 41.3 45.4 32.9 53.1 46 37.3
74.7 42.2 60.2 47.6 12.2 25.7 16 31.1 29.6 35.7 39 45.2 33.9 34.3
64.9 29 40.5 38.3 10.6 27.7 24 14.9 15.1 22.1 19.5 21.7 21.2 20.7
35.7 17.9 20.5 20.9 5.8 27.7 48 2.5 4.2 3.8 5.9 5.4 3.8 7.3 5.1 3.9
3 4.5 1.4 32.1 72 0 0.3 1 1 0.3 1.1 2.7 0.3 0 0 0.7 0.8 124.7
Parameter AUC.sub.0-12 h 731.2 625.0 582.6 504.3 711.6 698.5 535.4
683.5 509.8 793.2 637.5 101.1 15.9 (ng h/mL) AUC.sub.last 1270 1230
1343 1269 1568 1436 1354 1920 1101 1520 1401.1 229.0 16.3 (ng h/mL)
AUC.sub.inf 1301 1234 1358 1286 1571 1454 1418 1923 1164 1557
1426.6 218.9 15.3 (ng h/mL) C.sub.max 95.3 75.2 71.5 62 75.9 76.5
56.8 74.7 58.8 85.8 73.3 11.9 16.3 (ng/mL) T.sub.max (hours) 3 5 5
5 5 4 6 12 6 6 5.70 2.41 42.2 T.sub.1/2 (hours) 8.65 9.01 10.57
11.58 8.37 10.78 16.4 7.25 11.05 8.54 10.22 2.59 25.3
TABLE-US-00075 TABLE 70 Individual subject d-amphetamine
concentrations and pharmacokinetic parameters following oral
administration of a 30 mg dose of Dexedrine Spansule .RTM.
(equivalent to 75 mg dose of L-lysine-d-amphetamine based on
amphetamine base content) to humans Subject Subject Subject Subject
Subject Subject Subject Subject Subject Subject 102 103 105 107 110
112 113 116 117 120 Mean SD CV % Time Hours 0 0 0 0 0 0 0 0 0 0 0 0
0 0 0.5 1.2 2.68 1.37 1.4 1.16 2.36 6.75 2.63 4.95 3.43 2.8 1.8
65.5 1 14.8 26.5 16.7 21.4 25.2 12.7 33.1 22.3 26 21.5 22.0 6.1
27.8 1.5 24.2 36.9 23.2 28.5 37.2 21.3 42.4 29.2 33.7 39.2 31.6 7.3
23.2 2 28.6 43.4 27.3 34.6 38.5 27.6 46.2 31.3 38.5 42 35.8 6.9
19.4 3 27.4 37.3 30.6 40.1 41.7 30.9 52 36.5 42.9 60.1 40.0 10.0
25.2 4 27.1 44.1 33.5 48.7 45.2 34.7 49.1 40.7 42.4 53.2 41.9 8.1
19.2 5 35.1 53 40.2 43.4 46.5 42.4 58.1 47 52.1 68.7 48.7 9.7 20.0
6 33.8 58.5 40.2 46.5 43.5 37.5 56.2 40 51 63 47.0 9.8 20.8 7 37.2
50.7 31.2 41.4 44.9 42 57.8 43.6 51.6 65.7 46.6 10.1 21.7 8 35.9
54.3 34.9 45 45 36 58.7 41.8 53.9 59.2 46.5 9.5 20.4 10 33.1 49.1
34.3 35.5 45 37 51.4 38.9 46.3 60.1 43.1 8.8 20.4 12 34 51 28.6
34.1 40.8 32.6 51.6 37.7 38.1 50.9 39.9 8.4 21.1 16 30.2 40.8 25.2
28 33 25.8 41 26.8 29.6 44.9 32.5 7.1 22.0 24 20.5 27.8 18.2 19.5
17.1 17.8 22.5 19.1 15.5 27.3 20.5 4.2 20.3 48 3.83 6.89 3.7 5.11
2.56 4.31 6.51 4.43 2.77 5.47 4.6 1.4 31.8 72 0.715 1.63 1 1.7 0
0.622 1.29 1.22 0 1.31 0.9 0.6 64.0 Parameter AUC.sub.0-12 h 356.2
539.8 366.4 444.3 480.8 387.0 591.4 436.5 512.8 634.2 474.9 94.7
19.9 (ng h/mL) AUC.sub.last (ng h/mL) 1033 1517 966 1135 1065 1003
1473 1100 1048 1589 1193 236 19.8 AUC.sub.inf (ng h/mL) 1043 1544
983.5 1168 1097 1013 1495 1121 1085 1610 1216 238 19.5 C.sub.max
(ng/mL) 37.2 58.5 40.2 48.7 46.5 42.4 58.7 47 53.9 68.7 50.18 9.74
19.4 T.sub.max (hours) 7 6 5 4 5 5 8 5 8 5 5.80 1.40 24.1 T.sub.1/2
(hours) 9.92 11.74 12.07 13.8 8.7 10.76 11.47 12.23 9.36 10.92
11.10 1.50 13.6
TABLE-US-00076 TABLE 71 Pharmacokinetic parameters of amphetamine
following oral administration of L-lysine-d-amphetamine, Adderall
XR .RTM. or Dexedrine Spansule .RTM.. Drug L-lysine- L-lysine- d-
d- amphetamine amphetamine Adderall Dexedrine Parameter 25 mg
Percent.sup.1 75 mg Percent.sup.1 XR .RTM. Percent.sup.1 Spansule
.RTM. Percent.sup.1 AUC.sub.0-12 h 205.4 33.6 611.5 100 637.5 104
474.9 78 (ng h/mL) AUC.sub.last (ng h/mL) 396.7 31.5 1237 100
1401.1 113 1193 96 AUC.sub.inf (ng h/mL) 415 .+-. 80.3 32.9 1260
.+-. 192 100 1429 .+-. 223 113 1217 .+-. 237 97 C.sub.max (ng/mL)
25.0 .+-. 5.57 33.8 74.0 .+-. 12.9 100 73.3 .+-. 11.9 99 50.2 .+-.
9.74 68 T.sub.max (hours) 3.10 .+-. 0.88 79.5 3.90 .+-. 0.99 100
5.70 .+-. 2.41 146 5.8 .+-. 1.40 149 T.sub.1/2 (hours) 9.66 .+-.
1.45 94 10.3 .+-. 1.66 100 10.2 .+-. 2.62 99 11.1 .+-. 1.48 108
.sup.1Percent relative to L-lysine-d-amphetamine 75 mg dose
Example 34
Clinical Pharmacokinetic Evaluation and Oral Bioavailability of
L-lysine-d-amphetamine Dimesylate
[0383] In pediatric patients (6-12 yrs) with ADHD, the T.sub.max of
d-amphetamine was approximately 3.5 hours following single-dose
oral administration of L-lysine-d-amphetamine dimesylate either 30
mg, 50 mg, or 70 mg after a 8-hour overnight fast. See FIG. 65. The
T.sub.max of L-lysine-d-amphetamine dimesylate was approximately 1
hour. Linear pharmacokinetics of d-amphetamine after single-dose
oral administration of L-lysine-d-amphetamine dimesylate was
established over the dose range of 30 mg to 70 mg in children.
TABLE-US-00077 TABLE 72 Pharmacokinetic parameters of d-amphetamine
and L-lysine-d-amphetamine dimesylate C.sub.max T.sub.max AUC
t.sub.1/2 Dose (ng/mL) (h) (ng h/mL) (h) d-amphetamine 30 mg 53.2
.+-. 9.62 3.41 .+-. 1.09 845 .+-. 117 8.90 .+-. 1.33 50 mg 93.3
.+-. 18.2 3.58 .+-. 1.18 1510 .+-. 242 8.61 .+-. 1.04 70 mg 134
.+-. 26.1 3.46 .+-. 1.34 2157 .+-. 383 8.64 .+-. 1.32 Intact
L-lysine-d-amphetamine dimesylate 30 mg 21.9 0.97 27.9 50 mg 46.0
0.98 57.9 70 mg 89.5 1.07 108.9
[0384] There is no unexpected accumulation of d-amphetamine at
steady state in children with ADHD and no accumulation of
L-lysine-d-amphetamine dimesylate after once-daily dosing for 7
consecutive days.
[0385] Food does not affect the extent of absorption of
d-amphetamine in healthy adults after single-dose oral
administration of 70 mg of L-lysine-d-amphetamine dimesylate
capsules but delays T.sub.max by approximately 1 hour (from 3.78
hrs at fasted state to 4.72 hrs after a high fat meal). After an
8-hour fast, the extent of absorption of d-amphetamine following
oral administration of L-lysine-d-amphetamine dimesylate in
solution and as intact capsules was equivalent.
[0386] There were no apparent differences between males and females
in exposure as measured by dose-normalized C.sub.max and AUC
although the range of values in children was higher than that in
adults. This is a consequence of the significant correlation
between dose-normalized C.sub.max and AUC and body weight and thus
the differences are due to the higher doses in mg/kg administered
to children. There were no apparent differences in t.sub.1/2
between male and female subjects nor were there any apparent
relationships between t.sub.1/2 and either age or body weight.
[0387] Exemplary results of clinical pharmacokinetic evaluation are
presented in FIG. 66 (AUC), FIG. 67 (C.sub.max), and FIG. 68
(T.sub.max).
Example 35
Efficacy of L-lysine-d-amphetamine Dimesylate in Pediatric Clinical
Trials
[0388] The efficacy of L-lysine-d-amphetamine dimesylate was
established in a double-blind, randomized, placebo-controlled,
parallel-group study conducted in children aged 6-12 (N=290) who
met DSM-IV criteria for ADHD (either the combined type or the
hyperactive-impulsive type). Patients were randomized to fixed dose
treatment groups receiving final doses of 30, 50, or 70 mg of
L-lysine-d-amphetamine dimesylate or placebo once daily in the
morning for four weeks. For patients randomized to 50 and 70 mg
L-lysine-d-amphetamine dimesylate, dosage was increased by forced
titration. Significant improvements in the signs and symptoms of
ADHD, as rated by investigators (ADHD Rating Scale; ADHD-RS) and
parents (Connor's Parent Rating Scale; CPRS), were demonstrated for
all L-lysine-d-amphetamine dimesylate doses compared to placebo,
for all four weeks, including the first week of treatment, when all
L-lysine-d-amphetamine dimesylate patients were receiving a dose of
30 mg/day. Additional dose-responsive improvement was demonstrated
in the 50 and 70 mg groups, respectively. L-lysine-d-amphetamine
dimesylate-treated patients showed significant improvements, as
measured by CPRS scores, in the morning (.about.10 am), afternoon
(.about.2 pm), and evening (.about.6 pm) compared with
placebo-treated patients, demonstrating effectiveness throughout
the day. The results of the primary efficacy analysis, ADHD-RS
total score change from baseline to endpoint for the ITT
population, are shown in FIG. 69.
[0389] Efficacy was also measured by the SKAMP score. A total of 52
children ages 6 to 12 who met DSM-IV criteria for ADHD (either the
combined type or the hyperactive-impulsive type) were enrolled in a
double-blind, randomized, placebo-controlled crossover study.
Patients were randomized to receive fixed and optimal doses of
L-lysine-d-amphetamine (30, 50, 70 mg), Adderall XR.RTM. (10, 20,
or 30 mg), or placebo once daily in the morning for 1 week each
treatment. The primary efficacy endpoint in this study was
SKAMP-Deportment score (Swanson, Kotkin, Agler, M. Flynn and Pelham
rating scale). Both L-lysine-d-amphetamine and Adderall XR.RTM.
were highly effective compared to placebo. The significant effects
of L-lysine-d-amphetamine occurred within 2 hours post morning dose
and continued throughout the last assessment time point, 12 hours
post morning dose, compared to placebo, yielding a 12-hour duration
of action. See FIG. 70.
Example 36
Abuse Liability of Intravenous L-lysine-d-amphetamine
[0390] L-lysine-d-amphetamine 50 mg, d-amphetamine 20 mg, and
placebo were given intravenously over 2 minutes at 48 hour
intervals to 9 stimulant abusers in a double blind crossover design
to assess abuse liability. Drugs were given according to 3.times.3
balanced latin squares. Each dosing day, vital sign measures and
subjective and behavioral effects were assessed with questionnaires
before dosing and at 0.5, 1, 1.5, 2, 3, 4, 5, 6, 9, 12, 16 and 24
hours after dosing. At these times and at 5 minutes, a blood sample
(5 ml) was taken for d-amphetamine levels.
[0391] For d-amphetamine, mean peak plasma level of 77.7 ng/ml of
d-amphetamine occurred at 5 minutes and then rapidly subsided.
Administration of d-amphetamine produced expected
d-amphetamine-like effects with mean peak responses at 15 minutes.
The mean maximum response to d-amphetamine on the primary variable
of Subject Liking VAS was significantly greater than placebo
(p=0.01).
[0392] For L-lysine-d-amphetamine, mean peak plasma level of 33.8
ng/ml of d-amphetamine occurred at 3 hours and remained at this
level through the 4 hour observation. L-lysine-d-amphetamine
produced d-amphetamine-like subjective, behavioral, and vital sign
effects with mean peak responses at 1 to 3 hours. For the primary
variable of Subject Liking VAS, the response was not greater than
placebo (p=0.29). Changes in blood pressure following
L-lysine-d-amphetamine were significant.
[0393] At the end of the study, subjects were asked which treatment
they would take again. Six subjects chose d-amphetamine 20 mg, two
subjects chose none of the treatments, and one subject chose
L-lysine-d-amphetamine 50 mg. In summary, L-lysine-d-amphetamine 50
mg did not produce euphoria or amphetamine-like subjective effects
although there were late occurring blood pressure increases. The
findings suggest that L-lysine-d-amphetamine itself is inactive.
After 1 to 2 hours, L-lysine-d-amphetamine is converted to
d-amphetamine. Taken intravenously, L-lysine-d-amphetamine has
significantly less abuse potential than immediate release
d-amphetamine containing an equal amount of d-amphetamine base.
Example 37
Preliminary Estimates of Decreased Abuse Liability with
L-lysine-d-amphetamine Vs. d-amphetamine in Healthy Adults with a
History of Stimulant Abuse
[0394] This randomized, single-center, single-blind,
dose-escalation study used pharmacokinetic parameters to obtain
preliminary estimates of abuse liability for L-lysine-d-amphetamine
(30-150 mg) vs. d-amphetamine sulfate (40 mg) and placebo in
healthy adults meeting DSM-IV criteria for stimulant abuse.
Subjects were divided into 3 cohorts of 4 patients each; all
received single doses of L-lysine-d-amphetamine at a minimum
interval of 48 hours, with d-amphetamine sulfate (40 mg) and
placebo randomly dispersed. Cohort 1 was administered
L-lysine-d-amphetamine doses of 30, 50, 70, 100 mg; cohort 2
received 50, 70, 100, 130 mg doses; and cohort 3 received 70, 100,
130, and 150 mg doses.
[0395] AUC.sub.last d-amphetamine over the first 4 hours was
substantially lower with 100 mg L-lysine-d-amphetamine (165.3-213.1
ng/mL) vs. 40 mg d-amphetamine (245.5-316.8 ng/mL). C.sub.max and
AUC.sub.last increased with dose for 30-130 mg
L-lysine-d-amphetamine, attenuating between the 130 mg and 150 mg
dose. T.sub.max ranged from 3.78-4.25 h with L-lysine-d-amphetamine
vs. d-amphetamine sulfate (1.88-2.74 h). The half-life of
L-lysine-d-amphetamine (range, 0.44-0.76 h) indicated rapid
clearance. Adverse reactions were mild in severity with no
significant changes in vital signs or ECG parameters.
L-lysine-d-amphetamine had a slower release of d-amphetamine
compared with d-amphetamine sulfate. At doses as high as 150 mg,
there appears to be an attenuation of the maximum concentration,
suggesting higher doses of L-lysine-d-amphetamine will not lead to
further increases in C.sub.max and AUC.sub.last. These results
suggest a drug profile consistent with reduced abuse liability.
Example 38
Preparation of L-lysine-d-amphetamine Dimesylate
[0396] L-lysine-d-amphetamine dimesylate was prepared according to
the procedure shown in FIG. 2 and as described in more detail
below.
[0397] L-Lysine monohydrochloride (1 wt=1 eq) and water (6 vol)
were added to a clean vessel and then heated to .about.35.degree.
C. The pH of the mixture was adjusted to .about.11-11.5 using 50%
sodium hydroxide solution. Di-tert-butyl dicarbonate and 50% sodium
hydroxide solution were charged to the vessel at a rate such that
the temperature was maintained at .about.30-50.degree. C. and
pH>8.
[0398] The reaction mixture was then heated to .about.50.degree. C.
and the temperature was maintained for 2 hr., or until reaction
completion. The mixture was cooled to .about.20-25.degree. C.,
followed by dilution with isopropyl acetate (IPAc) (6 vol.). The pH
of the mixture was adjusted to .about.2.0-2.5 using concentrated
hydrochloric acid. Then, the mixture was agitated for 15 min.
followed by phase separation. The organic phase was washed with
brine (25% wt/wt, 1.34 vol.).
[0399] The mixture containing the non-isolated intermediate
Boc-L-Lys(Boc)-OH was charged on top of a N-hydroxysuccinimide
solution (1.1 eq.) in a clean vessel. The original vessel was then
rinsed with IPAc (2.5 vol.) and the washings were combined with the
mixture. The temperature of the mixture was adjusted to
.about.50.degree. C. A solution of N,N'-Dicyclohexyl carbodiimide
(1.05 eq.) and IPAc (1.2 vol.) heated to .about.30.degree. C. was
then added. The temperature of the reaction mixture was maintained
at .about.50.degree. C. The mixture was then agitated for 2 hr., or
until reaction completion. The mixture was cooled to
.about.20-25.degree. C. followed by filtration to remove
N,N'-dicyclohexylurea.
[0400] Following filtration, the filter-cake was rinsed with IPAc
(4 vol.). The filtrate (IPAc solution) was washed twice with
saturated sodium bicarbonate solution (1.3 vol. per wash). The IPAc
solution was washed with water (1.2 vol.) and then brine (25%
wt/wt, 1.34 vol.). The reaction mixture was concentrated to 12 vol.
by vacuum distillation below 50.degree. C. to give the non-isolated
intermediate Boc-L-Lys(Boc)-OSu.
[0401] d-amphetamine (1.1 eq.), 4-methylmorpholine (1.1 eq) and
IPAc (12 vol.) were charged into a clean vessel. The solution of
Boc-L-Lys(Boc)-OSu in IPAc was then charged into the same vessel at
a rate such that the temperature remained below 30.degree. C. The
reaction mixture was agitated for 2 hr. at .about.20-30.degree. C.,
or until reaction completion.
[0402] A quench solution was prepared by mixing sodium chloride
(4.5 wt), glacial acetic acid (0.3 wt) and water (18 vol), and
agitating it until dissolution was achieved. One half of the quench
solution was added to the reaction mixture. The reaction mixture
was agitated for 30 min., followed by phase separation. The
remaining quench solution was added. The reaction mixture was
washed with aqueous sodium bicarbonate solution (8% wt/wt, 11.5
vol.), followed by brine (7% wt/wt, 11.5 vol.). .about.6.25 vol.
was removed from the reaction mixture by vacuum distillation at
.about.55-65.degree. C. Isopropyl alcohol (IPA) (21 vol.) was
added. The mixture was concentrated to 6.25 vol. by vacuum
distillation at .about.55-65.degree. C. IPA (2.5 vol.) was again
added, and the reaction mixture was filtered through a 1.2 mm
filter. Methane sulfonic acid (2.05 eq.) and water (0.6 vol.) were
added to the filtrate and the mixture was heated to
.about.60-70.degree. C. for 6 hr. at a partial vacuum (.about.550
mmHg). When complete, pre-heated (60-70.degree. C.) IPAc (7.7 vol.)
was added to the mixture while the temperature was maintained at
60-70.degree. C. The mixture was then agitated at 60-70.degree. C.
for 1-2 hrs., followed by cooling to 20-25.degree. C. by lowering
the jacket temperature .about.10.degree. C./hour. The temperature
was adjusted to 50-60.degree. C. for 2-3 hours. The mixture was
then cooled to 20-25.degree. C. by lowering the jacket temperature
.about.10.degree. C./hour and aged at 20-25.degree. C. for at least
16 hr.
[0403] The product was collected by filtration. The filter-cake was
washed twice with a mixture of IPA and IPAc (2:1, 10.5 vol per
wash), and dried at .about.50-60.degree. C. in vacuo. The dried
material was screened through a screener/magnet (20-mesh) to yield
the final crystalline product.
[0404] Optical micrographs and SEM (scanning electron microscopy)
images of the crystals prepared are shown in FIGS. 71 and 72,
respectively.
[0405] Particle size determination provided d.sub.10, d.sub.50 and
d.sub.90 values of 5 .mu.m, 26 .mu.m, and 57 .mu.m,
respectively.
Example 39
Preparation of L-lysine-d-amphetamine Dimesylate
[0406] L-lysine-d-amphetamine dimesylate was prepared according to
the procedure shown in FIG. 87 and as described in more detail
below.
[0407] A. Preparation of d-amphetamine Free Base
[0408] D-amphetamine sulfate USP (1 eq.) and water (.about.138 eq.)
were added to a vessel and agitated to dissolve the solids. The pH
of the mixture was adjusted to 10.5-11.5 using 50% sodium hydroxide
solution. The mixture was diluted with IPAc (.about.13.3 eq.). The
mixture was agitated for 30-45 min., followed by phase separation.
IPAc (.about.2.7 eq.) was then added to the aqueous phase. The
mixture was agitated for 30-45 min. and the phases were then again
separated. The organic phase was combined with the organic phase
from the previous phase separation. If the water content of the
combined dehydrated organic phase was >4.0%, water (.about.17.3
eq.) and sodium chloride (.about.1.8 eq.) were added to the organic
phase; the mixture was agitated for 30-45 min., the phases
separated, and the dehydrated organic phase was then used for
further processing.
[0409] B. Preparation of Boc-L-Lys(Boc)-OSu
[0410] L-Lysine monohydrochloride (1 wt.=1 eq.) and water (6 vol.)
were heated to .about.35.degree. C. in a clean vessel. The pH of
the mixture was adjusted to approximately 11-11.5 using 50% sodium
hydroxide solution. Di-tert-butyl dicarbonate and 50% sodium
hydroxide solution were then charged to the vessel at a rate
allowing to maintain the mixture temperature at approximately
30-60.degree. C. and the pH between 8.5-10. The reaction mixture
was heated to approximately 50.degree. C. and the temperature was
maintained for 2 hr., or until reaction completion. The mixture was
subsequently cooled to approximately 20-25.degree. C., and diluted
with IPAc (6 vol.). The pH was then adjusted to approximately
2.0-2.5 using concentrated hydrochloric acid. The mixture was
agitated for 30-45 min. and phases were then separated. The organic
phase was washed with brine (25% wt/wt, 1.34 vol.).
N-hydroxysuccinimide (1.1 eq.) was charged into a clean vessel. The
mixture containing the non-isolated intermediate Boc-L-Lys(Boc)-OH
was charged on top of the N-hydroxysuccinimide. The original vessel
was rinsed with IPAc (2.5 vol.) and the washings were combined with
the mixture. After which, the mixture temperature was adjusted to
approximately 50.degree. C.
[0411] N,N'-Dicyclohexyl carbodiimide (1.05 eq.) was charged to the
mixture while maintaining the temperature at approximately
50.degree. C. IPAc (3.7 vol.) was charged on top of the mixture at
a rate allowing to maintain the mixture temperature at
approximately 50.degree. C. The mixture was agitated for at least 2
hr., or until reaction completion. The mixture was cooled to
approximately 20-25.degree. C. The mixture was filtered to remove
N,N'-dicyclohexylurea. The filter-cake was rinsed with IPAc (5.9
vol.). The filtrate (IPAc solution) was washed with saturated
sodium bicarbonate solution (2.6 vol.). The IPAc solution was
washed with water (1.2 vol.) and then brine (25% wt/wt, 1.34
vol.).
[0412] The mixture was then concentrated to 25-65% of the starting
volume via vacuum distillation below 35.degree. C. to give the
non-isolated intermediate Boc-L-Lys(Boc)-Osu. The mixture was
filtered through a filter dressed with diatomaceous earth
impregnated pads.
[0413] C. Preparation of L-lysine-d-amphetamine Dimesylate
[0414] Freebased d-amphetamine (1.1 eq.), 4-methylmorpholine (1.085
eq.) and IPAc (12 vol.) were added to a clean vessel. The solution
of Boc-L-Lys(Boc)-OSu in IPAc was added at a rate such that the
temperature of the reaction mixture remained below 30.degree. C.
The mixture was agitated for 1-2 hr. at approximately 20-30.degree.
C., or until reaction completion.
[0415] A quench solution was prepared by mixing sodium chloride
(2.25 wt.), glacial acetic acid (0.15 wt) and water (9 vol.), and
agitating the mixture until dissolution was achieved. One half of
the quench solution was added to the reaction mixture, the mixture
was agitated for 30-45 min., and the phases were separated. The
remaining quench solution was added followed by agitation for 30-45
min. and phase separation.
[0416] The IPAc solution was washed with aqueous sodium bicarbonate
solution (8% wt/wt, 11.5 vol.) and then brine (7% wt/wt, 11.5
vol.). The mixture was concentrated to 25-40% of its starting
volume by vacuum distillation at .about.25-30.degree. C. IPA (21
vol.) was added and the mixture was again concentrated to 25-40% of
its starting volume by vacuum distillation at .about.25-30.degree.
C. IPA (2.5 vol.) was added, the mixture was filtered through a
0.45 mm filter, and rinsed with IPA (2.5 vol.).
[0417] Methane sulfonic acid (2.05 eq.) and water (0.6 vol.) were
added to the filtrate, and the mixture was heated to
.about.60-70.degree. C. for 8 hr. at a partial vacuum (.about.550
mmHg). IPAc (.about.12.1 eq.) was added to the reaction mixture
while maintaining its temperature at 60-70.degree. C. The reaction
mixture was stirred for 1-2 hrs. at 60-70.degree. C., and cooled to
.about.20-25.degree. C. at a rate of approximately 10.degree.
C./hr. The mixture was heated to 60-70.degree. C., stirred for 2-3
hrs., and cooled to approximately 20-25.degree. C. at a rate of
approximately 10.degree. C./hr. The slurry was stirred for 16-16.5
hrs at 20-25.degree. C. The product was filtered, washed twice with
a mixture of IPA and IPAc (2:1, 10.5 vol. per wash), and dried at
.about.65.+-.5.degree. C. under full vacuum. The dried material was
milled through a 16-20 mesh screen to yield the final crystalline
product.
[0418] Optical micrographs of the crystals prepared are in FIGS. 73
and 74. SEM images of the crystals prepared are shown in FIGS. 75
and 76.
[0419] Particle sizes, in terms of d.sub.10, d.sub.50 and d.sub.90
values, were determined to be 4.1 .mu.m, 19.5 .mu.m and 60.8 .mu.m,
respectively.
Example 40
Characterization of Crystalline L-lysine-d-amphetamine
Dimesylate
[0420] XRPD
[0421] The XRPD spectra of a sample of the crystalline
L-lysine-d-amphetamine dimesylate prepared according to example 39
is shown in FIG. 77. Peak locations (in degrees 2.THETA..+-.0.2,
0.1, 0.05, or 0.01.degree. C.) for the XRPD pattern in FIG. 77 are
provided in table 73, below.
TABLE-US-00078 TABLE 73 Peak locations 2.THETA. d space (.ANG.)
Intensity 4.5 .+-. 0.2 19.534 .+-. 0.441 100 9.0 .+-. 0.2 9.805
.+-. 0.110 19 12.0 .+-. 0.2 7.400 .+-. 0.062 24 15.7 .+-. 0.2 5.637
.+-. 0.036 33 16.3 .+-. 0.2 5.453 .+-. 0.034 6 17.2 .+-. 0.2 5.143
.+-. 0.030 11 17.5 .+-. 0.2 5.061 .+-. 0.029 3 17.8 .+-. 0.2 4.971
.+-. 0.028 9 18.1 .+-. 0.2 4.903 .+-. 0.027 13 18.5 .+-. 0.2 4.785
.+-. 0.026 4 18.9 .+-. 0.2 4.684 .+-. 0.025 29 19.2 .+-. 0.2 4.624
.+-. 0.024 18 19.8 .+-. 0.2 4.492 .+-. 0.023 8 20.5 .+-. 0.2 4.333
.+-. 0.021 12 20.8 .+-. 0.2 4.277 .+-. 0.020 6 21.6 .+-. 0.2 4.123
.+-. 0.019 3 22.8 .+-. 0.2 3.899 .+-. 0.017 31 23.3 .+-. 0.2 3.814
.+-. 0.016 11 23.5 .+-. 0.2 3.782 .+-. 0.016 5 23.9 .+-. 0.2 3.719
.+-. 0.015 14 24.1 .+-. 0.2 3.691 .+-. 0.015 25 24.6 .+-. 0.2 3.615
.+-. 0.015 7 25.2 .+-. 0.2 3.532 .+-. 0.014 3 25.5 .+-. 0.2 3.489
.+-. 0.013 7 26.5 .+-. 0.2 3.366 .+-. 0.013 2 27.3 .+-. 0.2 3.273
.+-. 0.012 9 27.8 .+-. 0.2 3.215 .+-. 0.011 8 28.9 .+-. 0.2 3.092
.+-. 0.011 2 29.7 .+-. 0.2 3.007 .+-. 0.010 5
[0422] DSC
[0423] DSC thermograms were generated for thirteen different
samples of L-lysine-d-amphetamine dimesylate, which were prepared
according to the procedure in example 38 or 39. Each sample
exhibited three endothermic events at approximately 94.degree. C.,
170.degree. C. and 194.degree. C. A representative DSC thermogram
is shown in FIG. 78.
[0424] FT-Raman Spectra
[0425] The FT-Raman spectra of L-lysine-d-amphetamine dimesylate
prepared according to the procedure in example 38 or 39 are shown
in FIGS. 79 and 80. This crystal form of L-lysine-d-amphetamine
dimesylate exhibits characteristic peaks at one or more of 1003,
1042, 2918, 2935, and 3055 cm.sup.-1. The peaks observed in the
FT-Raman spectra and their relative intensities are provided below
in tables 74 and 75.
TABLE-US-00079 TABLE 74 Peak list for the FT-Raman Spectrum in FIG.
79 Position Intensity Position Intensity 125 40.311 229 18.864 286
6.445 344 20.104 392 7.222 426 3.248 451 3.964 464 4.618 496 3.758
511 6.449 530 7.205 556 21.639 601 3.070 621 7.989 683 2.113 745
5.327 778 25.979 814 6.988 849 2.680 863 4.269 876 4.851 904 5.906
942 3.913 958 3.471 1003 44.089 1042 46.907 1085 8.540 1097 7.210
1158 9.285 1182 12.150 1208 26.723 1263 7.095 1273 6.033 1291 7.508
1304 7.078 1332 10.244 1342 12.031 1378 3.474 1420 10.636 1446
20.092 1500 3.111 1550 3.080 1585 12.398 1601 12.963 1607 17.523
1656 13.423 2656 2.107 273: 2.911 2785 2.876 2852 7.447 2872 10.363
2918 46.220 2935 81.701 2978: 24.925 3004 16.222 3017: 19.421 3042
14.820 3055 33.276 3319 1.417
TABLE-US-00080 TABLE 75 Peak list for the FT-Raman Spectrum in FIG.
80 Position Intensity Position Intensity 125 40.311 229 18.864 344
20.104 392 7.222 556 21.639 621 7.989 778 25.979 814 6.988 876
4.851 904 5.906 1003 44.089 1042 46.907 1085 8.540 1182 12.150 1208
26.723 1342 12.031 1446 20.092 1585 12.398 1607 17.523 1656 13.423
2918 46.220 2935 81.701 2978 24.925 3017 19.421 3055 33.276
[0426] Single Crystal Analysis
[0427] A single crystal analysis was performed as follows. The
l-lysine-d-amphetamine dimesylate was re-crystallized with a 1:1
acetonitrile/ethanol mixture and slurried at elevated temperature
in acetonitrile overnight to obtain crystals suitable single
crystal X-ray structure determination.
[0428] Single crystal data for the L-lysine-d-amphetamine
dimesylate crystals was determined with Mo K.sub..alpha. radiation
(.lamda.=0.71073 .ANG.) on a Nonius KappaCCD diffractometer
equipped with a graphite crystal, incident beam monochromator.
Refinements were performed on an LINUX PC using SHELX97.
[0429] Cell constants and an orientation matrix for data collection
were obtained from least-squares refinement using the setting
angles of 13580 reflections in the range
2.degree.<.theta.<24.degree.. The space group was determined
by the program XPREP. From the systematic presence of the following
conditions: 0k0 k; =2n, and from subsequent least-squares
refinement, the space group was determined to be P 2.sub.1 (no. 4).
This is a chiral space group.
[0430] The data were collected to a maximum 2.theta. value of
48.23.degree., at a temperature of 150.+-.1 K.
[0431] Frames were integrated with DENZO-SMN. A total of 13580
reflections were collected, of which 5474 were unique. Lorentz and
polarization corrections were applied to the data. The linear
absorption coefficient is 0.270 mm.sup.-1 for Mo K.sub..alpha.
radiation. An empirical absorption correction using SCALEPACK was
applied. Transmission coefficients ranged from 0.884 to 0.987.
Intensities of equivalent reflections were averaged. The agreement
factor for the averaging was 13% based on intensity.
[0432] The structure was solved by direct methods using SIR2004.
The remaining atoms were located in succeeding difference Fourier
syntheses. Hydrogen atoms were included in the refinement but
restrained to ride on the atom to which they are bonded. The
structure was refined in full-matrix least-squares by minimizing
the function:
.SIGMA.w(|F.sub.o|.sup.2-|F.sub.c|.sup.2).sup.2
[0433] The weight w is defined as
1/[(.sigma..sup.2(F.sub.o.sup.2)+(0.1193P).sup.2+(9.5997P)], where
P=(F.sub.o.sup.2+2F.sub.c.sup.2)/3.
[0434] Scattering factors were taken from the "International Tables
for Crystallography". Of the 5474 reflections used in the
refinements, only the reflections with
F.sub.o.sup.2>2.sigma.(F.sub.o.sup.2) were used in calculating
R. A total of 5089 reflections were used in the calculation. The
final cycle of refinement included 546 variable parameters and
converged (largest parameter shift was essentially equal to its
estimated standard deviation) with unweighted and weighted
agreement factors of:
R = F o - F c / F o = 0.091 R w = ( w ( F o 2 - F c 2 ) 2 / w ( F o
2 ) 2 ) = 0.228 ##EQU00001##
[0435] The standard deviation of an observation of unit weight was
1.119. The highest peak in the final difference Fourier had a
height of 0.68 e/.ANG..sup.3. The minimum negative peak had a
height of -0.47 e/.ANG..sup.3. The factor for the determination of
the absolute structure refined to 0.2(2).
[0436] The results of this analysis are shown in table 76
below.
TABLE-US-00081 TABLE 76 Indexed and Single Crystal lattice
Parameters for Lisdexamphetamine Dimesylate Single Crystal Data
Indexed Solution Form/Pattern Single Crystal Solution Dimesylate
Family and Monoclinic Monoclinic Space Group P2.sub.1 #4 P2.sub.1/a
#14 Z'/Z 2/4 1/4 a (.ANG.) 10.2509(6) 10.304 b (.ANG.) 11.2804(6)
11.355 c (.ANG.) 19.3534(12) 19.499 .alpha. (deg) 90 90 .beta.
(deg) 94.124(3) 93.65 .gamma. (deg) 90 90 Volume (.ANG..sup.3/cell)
2232.1(2) 2277 V/Z (.ANG..sup.3/asym. unit) 558 569.3 Density
(g/cm.sup.3).sup.a 1.359 1.329 Weight Fraction Solvent 0 0
(%).sup.a
[0437] ORTEP and Packing Diagrams
[0438] ORTEP diagrams were prepared using the ORTEP III program
within the PLATON software package. These diagrams are shown in
FIGS. 81 and 82. Atoms are represented by 50% probability
anisotropic thermal ellipsoids.
[0439] Packing diagrams were prepared using CAMERON modeling
software. These diagrams are shown in FIGS. 83-85. Assessment of
chiral centers, void calculations and additional figures were
performed with the PLATON software package. Absolute configuration
was evaluated using the specification of molecular chirality rules.
Hydrogen bonding is represented as dashed lines.
[0440] XRPD Comparison
[0441] A comparison of a calculated XRPD pattern from the single
crystal parameters, and the XRPD pattern of lisdexamphetamine
dimesylate prepared according to Example 38 is shown in FIG.
86.
Example 41
Time Onset and Duration of Efficacy of L-lysine-d-amphetamine
Dimesylate in Pediatric Clinical Trials
[0442] A randomized, double-blind, multi-center,
placebo-controlled, two-way crossover, analog classroom study with
an open-label optimization phase, designed to assess the time of
onset, duration of efficacy, tolerability and safety of
l-lysine-d-amphetamine dimesylate (30, 50, and 70 mg) was
performed. The subjects of the study were children (males and
females) ages 6-12 years old (n=129) who satisfied DSM-IV-TR
criteria for a primary diagnosis of ADHD, combined or
hyperactive-impulse subtype. The study included four phases: (1)
Screening and washout; (2) open-label dose optimization; (3) analog
classroom sessions 1 and 2; and (4) 30-day safety follow-up.
Subjects were required to visit the clinic up to 8 times over a
7-10 week period.
[0443] Subjects were also required to have a baseline ADHD Rating
Scale IV (ADHD-RS-IV) score .gtoreq.28, age-appropriate
intellectual functioning, ability to complete the Permanent Product
Measure of Performance (PERMP) assessment, blood pressure
measurements within the 95th percentile for age, gender, and
height.
[0444] Key exclusion criteria included the presence of a comorbid
psychiatric condition with severe symptoms, conduct disorder, or
other medical condition that could, in the judgment of the
physician investigator, confound efficacy or safety assessments,
pose a risk to the subject, or prohibit study completion. Children
who had an adverse reaction (allergy, hypersensitivity,
intolerance) or who failed to respond to previous amphetamine
therapy were not allowed to participate in the study. Other
exclusion criteria were positive serum or urine pregnancy test,
history of substance abuse, weight <22.7 kg (50 lb), BMI
>98th percentile for age, seizure within the last 2 years, tic
or Tourette disorder, use of medication with effects on CNS
function or performance, and clinically significant laboratory and
ECG abnormalities. Children whose current ADHD medication provided
effective control of symptoms with acceptable tolerability were
also excluded.
[0445] Screening and Washout
[0446] Subjects were screened for approximately 3-weeks to
establish eligibility for study participation. Those subjects who
met eligibility requirements underwent medication washout, if
applicable. The length of the medication washout (if applicable)
was no longer than 7 days depending upon the half-life of the
subject's current ADHD medication.
[0447] Open-Label Dose Optimization
[0448] Following screening and washout, eligible subjects entered
the open-label dose optimization phase, during which they received
1 capsule of l-lysine-d-amphetamine dimesylate in an openlabel
manner and were evaluated for efficacy and tolerability to that
dosage approximately 7 days later. Dosage was initiated at 30 mg/d
l-lysine-d-amphetamine dimesylate and adjusted to the next
available dose, until optimal dose was reached. Optimal dose was
defined as the dose that produced a reduction in ADHD-RS-IV score
.gtoreq.30% and CGI-I score of 1 or 2 and had tolerable side
effects. Tolerability was determined by the investigator, based on
review of AEs and clinical judgement. Once reached, the optimal
dose was maintained for the remainder of the dose optimization
phase and was used for the double-blind treatment sequence
phase.
[0449] Dose increase was permitted if the current dose was well
tolerated, produced a reduction in ADHD-RS-IV score .gtoreq.30% and
CGI-I score of 1 or 2, and if the next available dose would, in the
opinion of the clinician, provide additional symptom reduction.
Dose reduction was permitted if tolerability to the current dose
was unacceptable; only 1 dose reduction was allowed. Subjects were
discontinued if they were unable to tolerate l-lysine-d-amphetamine
dimesylate or had not reached their optimal dose by visit 4/week 4.
The dose dispensed at visit 3 was used at visit 4 and the during
the double-blind treatment sequence period. During visit 4, a
practice analog classroom session was conducted, to familiarize
subjects with classroom schedules and procedures. Three to 5
practice PERMP tests were also given during the practice
session.
[0450] Double Blind Crossover Treatment with Analog Classroom
Sessions 1 and 2
[0451] Following completion of the open-label dose optimization
period and successful titration to an optimal dose of
l-lysine-d-amphetamine dimesylate, subjects entered the 2 week
double blind treatment portion of the study. Subjects were
randomized in a 1:1 ratio to 1 of 2 treatment sequences: daily
l-lysine-d-amphetamine dimesylate treatment (at the optimized dose)
for one week followed by daily placebo treatment (administered as
matched capsules identical in appearance to l-lysine-d-amphetamine
dimesylate capsules) treatment for one week, or vice versa.
[0452] For the first 6 days of each week during the double-blind
treatment phase, the study drug was administered by the parent. On
the last day of each week, the daily dose was administered by study
staff in the analog classroom, where efficacy and safety
assessments were also performed.
[0453] Each analog classroom had 10 to 18 subjects, with a maximum
of 16 from the same cohort. Subjects arrived at 6 AM, were
administered the SKAMP and PERMP at 0.5 hours predose (6:30 AM),
and received their randomized treatment (7 AM). SKAMP and PERMP
were then administered in the analog classroom at 1.5, 2.5, 5, 7.5,
10, 12 and 13 hours postdose. Subjects then departed at
approximately 8:30 PM.
[0454] Primary Efficacy
[0455] Efficacy was measured by the SKAMP score (teacher rating
scale) to evaluate the behavioral effects of l-lysine-d-amphetamine
dimesylate compared to placebo, under controlled conditions,
measured at multiple time points throughout the day. The SKAMP was
designed for independent observers to rate 13 items representing
two factors of classroom behavior--attention and deportment. A
detailed description of SKAMP can be found elsewhere (Wigal S B,
Gupta S, Guinta D, Swanson J M. Reliability and validity of the
SKAMP rating scale in a laboratory school setting. Psychopharmacol
Bull. 1998; 34:47-53).
[0456] In this study, a single, independent, trained observer rated
each subject on 13 items, using a 7-point impairment scale
(0=normal, 6=maximal impairment). Onset and duration of efficacy
were determined through SKAMP deportment assessments at 0.5 hours
predose and 1.5, 2.5, 5, 7.5, 10, 12 and 13 hours postdose.
[0457] The primary efficacy measure was the onset of action
measured by the mean SKAMP-DS score. A mixed linear model was used
to analyze the mean SKAMP-DS score as well as the SKAMP-DS scores
for each time point. In this model, the fixed effects were
sequence, period, and treatment, while the random effect was
subject-within-sequence. Raw means and effect sizes, least-squares
(LS) means and effect sizes, differences in LS means and 95%
confidence interval for the difference between treatment groups, p
values, and model result were calculated for each postdose
timepoint and for mean scores for the intent to treat (ITT)
population, i.e., the population which consisted of all randomized
subjects with at least 1 postrandomization measurement of the
primary efficacy variable (mean SKAMP deportment score over the
course of a day) available for analysis.
[0458] Secondary Efficacy
[0459] Secondary efficacy measures included the SKAMP subscales of
attention, quality of work, and compliance, SKAMP total score,
PERMP-A and -C, ADHD-RS-IV and the Clinical Global Impressions
(CGI). The PERMP is a 5-page math test consisting of 80 problems
each (total of 400 problems) and was used to evaluate efficacy and
to determine time of onset. Subjects were instructed to work at
their desks and to complete as many problems as possible in 10
minutes. Academic performance was evaluated using two scores:
PERMP-A (the number of problems attempted) and PERMP-C (the number
of problems correct). The appropriate level of difficulty for each
student was determined based on results of a math pre-test
administered at screening. The PERMP was completed during analog
classroom sessions at the same time points as the SKAMP scale. To
avoid taking the same test more than once during the study,
subjects were given a different version of the test at each
assessment time point.
[0460] The clinician completed ADHD-RS-IV was also used as a
secondary efficacy measure. The ADHD-RS-IV was administered at each
visit, beginning with Screening, and at baseline and each visit, to
capture the ADHD symptoms within each study week. The ADHD-RS-IV
consists of 18 items designed to reflect current symptomatology of
ADHD based on DSM-IV-TR.TM. criteria. The 18 items are grouped into
2 subscales (hyperactivity/impulsivity and inattentiveness). Each
item is scored on a scale of 0 (no symptoms) to 3 (severe
symptoms), yielding a total score of 0 to 54.
[0461] In addition to the ADHD-RS-IV, the Clinical Global
Impressions (CGI) Scale was used as a secondary efficacy measure.
The CGI rating scale permits a global evaluation of the subject's
improvement over time. At Baseline, a CGI-S assessment was
performed, in which the Investigator rates the severity of a
subject's condition on an 8-point scale ranging from 0 (not
assessed) to 7 (among the most extremely ill).
[0462] Responses for the CGI-I were dichotomized such that "very
much improved" (CGI-I score of 1) and "much improved" (CGI-I score
of 2) were combined into one category ("improved"), and the
remaining responses were combined into the other category ("not
improved").
[0463] The mixed linear model used to analyze scores for the
SKAMP-DS onset of action was also used to analyze scores for the
secondary efficacy measures.
[0464] Statistical Analysis
[0465] Determination of sample size was based on analysis of SKAMP
deportment scores from a previous crossover study scale 10.
Assuming a standard deviation SD) of 0.9491 (the maximum SD
reported in the previous crossover study), 96 subjects (48 subjects
in each treatment sequence) would need to complete the study to
detect a difference of 0.50 in mean SKAMP deportment scores between
placebo and l-lysine-d-amphetamine dimesylate at 95% power (P=0.05,
2-sided). However, 128 subjects were planned for enrollment, since
25% of subjects were predicted to discontinue during the study. All
statistical tests were 2-sided and performed at the 0.05
significance level.
[0466] Efficacy Outcomes.
[0467] As stated above, efficacy was demonstrated at each post-dose
timepoint assessed, starting at 1.5 hrs. postdose, and continuing
through the last (13.0 hrs. postdose). The primary efficacy results
are given in FIG. 88. At all timepoints, L-lysine-d-amphetamine
dimesylate had significantly lower LS mean SKAMP deportment scores
(DS) compared with placebo (P.ltoreq.0.01).
[0468] Secondary efficacy measurements were also demonstrated at
each postdose timepoint. As with the primary measurements,
L-lysine-d-amphetamine dimesylate had significantly lower LS mean
SKAMP attention scores compared with placebo, at all timepoints
(P<0.0001). These results are given in FIG. 89.
[0469] The results from the PERMP-A tests are given in FIG. 90.
Efficacy was demonstrated at each post-dose timepoint assessed,
starting at 1.5 hrs. postdose, and continuing through the last
(13.0 hrs. postdose). At all timepoints, L-lysine-d-amphetamine
dimesylate had significantly higher PERMP-A scores compared with
placebo (P.ltoreq.0.0001).
[0470] The results from the PERMP-C tests are given in FIG. 91.
Efficacy was demonstrated at each post-dose timepoint assessed,
starting at 1.5 hrs. postdose, and continuing through the last
(13.0 hrs. postdose). At all timepoints, L-lysine-d-amphetamine
dimesylate had significantly higher PERMP-A scores compared with
placebo (P.ltoreq.0.0001). As FIGS. 88-91 indicate,
L-lysine-d-amphetamine dimesylate has a duration of action (as
measured by efficacy) for at least 13 hours.
[0471] An analysis of the change from Baseline in ADHD-RS-IV Total
scores, Inattention subscale scores, and Hyperactivity/Impulsivity
subscale scores for the Cross-Over phase (Visit 5/6) is presented
for the ITT population in table 77 below. For all three ADHD-RS-IV
measures, l-lysine-d-amphetamine dimesylate treatment resulted in a
larger reduction from Baseline scores compared to placebo
treatment. For all three ADHD-RS-IV measures, change from Baseline
scores associated with l-lysine-d-amphetamine dimesylate treatment
were statistically significantly different (p<0.0001) from those
associated with placebo treatment.
TABLE-US-00082 TABLE 77 Analysis ADHD-RS-IV Change from Baseline
Total and Subscale Scores at Visit 5/6 by Treatment (ITT
population) Raw LS effect efect Diff. Raw mean (SE) size LS mean
(SE) size in LS 95% variable Drug placebo (SE) Drug placebo (SE)
means C.L. P value total score -25.78 -8.65 -1.33 -25.75 -8.66
-1.36 -17.1 -20.41, <0.0001 (1.22) (1.20) (0.16) (1.20) (1.20)
(0.16) -13.78 Inattention score 12.47 -4.12 -1.24 -12.45 -4.12
-1.35 -8.33 -9.96, <0.0001 (0.65) (0.61) (0.16) (0.62) (0.62)
(0.16) -6.71 Hyperactivity/impulsivity 13.31 -4.53 -1.28 13.30
-4.54 -1.31 -8.76 -10.52, <0.0001 score (0.64) (0.65) (0.16)
(0.64) (0.64) (0.16) -7.00
[0472] An analysis of CGI-I scores for the Cross-Over Phase (Visits
5 and 6) is presented in graphically in FIG. 92. A majority of the
113 subjects in the ITT population (71.7%) were improved while
receiving l-lysine-d-amphetamine dimesylate but not on placebo. Ten
subjects (8.8%) were not improved while receiving
l-lysine-d-amphetamine dimesylate but were improved on placebo. The
difference between l-lysine-d-amphetamine dimesylate and placebo
treatment was statistically significant (p<0.0001).
Example 42
Within and Between Subject Variability for Patients Administered
l-lysine-d-amphetamine Dimesylate
[0473] An open-label, five-period, five-treatment, dose escalating
study of the safety, tolerability, and pharmacokinetics of
l-lysine-d-amphetamine dimesylate administered as a single oral
dose was conducted. The study was conducted at a single center and
consisted of a Screening and five single Dosing Periods. During the
Screening Period (within 28 days prior to Dosing Period 1, Day 1
dosing), the eligibility of subjects were confirmed. Eligibility
was re-confirmed at check-in on Day 1. Eligible participants were
confined to the Clinical Research Center (CRC) after checking in on
Day 1 and remained in the CRC until approximately 96 hours post
Dosing Period 5 and after the scheduled discharge assessments and
procedures were completed. No washout period was given between
dosing periods. The total duration of the confinement for this
study was 22 days.
[0474] Subjects received a single dose of l-lysine-d-amphetamine
dimesylate on Day 1 of each Dosing Period and a total of five doses
over the course of the study. During each Dosing Period, serial
blood samples were collected up to 96 hours post dose. During this
time, subjects were required to fast from approximately 10:00 pm
the night before receiving study drug until four hours after taking
study drug. Subjects were discharged from the CRC at least 96 hours
after taking the last dose of study medication in Dosing Period 5
and once all discharge assessments and procedures were complete and
unremarkable. Approximately 7 days after the last dose of study
drug was administered, a follow-up was done to report any ongoing
concomitant medication(s), adverse events (AEs), and/or new AEs or
Serious Adverse Events (SAEs).
[0475] Pharmacokinetic Analysis:
[0476] Descriptive statistics (number of observations [N], mean,
standard deviation, coefficient of variation [CV], median, minimum,
and maximum) were used to summarize plasma concentration at each
planned sampling time point for each treatment. Plasma
pharmacokinetic parameters calculated from the concentrations were
also summarized by dose level using descriptive statistics.
[0477] Dose proportionality of the key pharmacokinetic parameter
AUC.sub.0-.infin. over the administered dose range for
d-amphetamine was based on the following power model:
log(parameter)=a+b*log(dose); where `a` is the intercept and `b` is
the slope.
[0478] The power model fitted for the log-transformed parameter
AUC.sub.0-.infin. with a mixed model included a random intercept.
The estimated mean slope (`b`) and 90% Confidence Intervals (CIs)
were constructed. Linearity was assessed based on a test of the
null hypothesis that the slope is equal to 1 versus the alternative
hypothesis that the slope is not equal to 1. The other
pharmacokinetic parameters were summarized as described above.
[0479] Pharmacokinetic Evaluations
[0480] On Day 1 of each dosing period, blood samples for
pharmacokinetic analysis were drawn pre-dose and, after a single
oral dose of l-lysine-d-amphetamine dimesylate, at 0.25, 0.5, 1,
1.5, 2, 4, 6, 8, 12, 16, 24, 36, 48, 72 and 96 hours post-dose. On
Day 5, after collecting the 96-hour post dose sample, subjects
progressed to the next dosing period. The 96-hour sample served as
the pre-dose sample for the next period. All pharmacokinetic data
are presented by dose (50, 100, 150, 200, 250 mg).
[0481] The mean d-amphetamine plasma concentration profiles for all
five lisdexamfetamine dimesylate dosing periods (50, 100, 150, 200,
250 mg) are shown in FIG. 93. Other pharmacokinetic parameters
measured in this study are given in table 78 below. Across all
doses administered, pharmacokinetic parameters were consistent and
well-behaved. The mean maximum observed concentration (C.sub.max)
and mean systemic exposure (AUC.sub.0-t, AUC.sub.0-.infin.)
increased proportionately with dose. The mean time to maximum
plasma concentration (t.sub.max) occurred at approximately 5 hours
and the mean terminal half life (t.sub.1/2) at approximately 11.5
hours.
TABLE-US-00083 TABLE 78 Mean Plasma Pharmacokinetic Parameters by
Dose for d-amphetamine after a Single Oral Dose of Lisdexamfetamine
Dimesylate Lisdexamfetamine Dimesylate Dose 50 mg 100 mg 150 mg 200
mg 250 mg C.sub.max (ng/mL) N 20 20 18 12 9 Mean 44.62 84.55 126.57
168.83 246.32 (SD) (9.31) .sup. 15.07) 29.47) .sup. 50.63) 100.81)
% CV 20.9 17.8 23.3 30 40.9 Median 43 84.79 117.61 175.77 230.49
(Min, Max) (33.58, 61.14) (66.96, 115.78) (89.75, 190.07) (43.26,
234.53) (148.2, 477.19) t.sub.max, hr N 20 20 18 12 9 Mean 4 4.5
4.9 5.7 5.8 (SD) (1.2) (0.9) (1.6) (0.8) (1.2) % CV 29.6 19.7 32
13.8 20.8 Median 4 4 4 6 6 (Min, Max) (1.5, 6) (4, 6) (2, 8) (4,
6.1) (4, 8) AUC.sub.0-t (ng h/mL).sup.a N 20 20 18 12 9 Mean 763.1
1485.1 2429.3 3265.5 5056.8 (SD) (190.4) (401.2) (727.2) (1207.7)
(1419.8) % CV 24.9 27 29.9 37 28.1 Median 732 1413.9 2237.9 3359.9
4924.3 (Min, Max) (546.8, 1382.9) (1105.9, 959.1) (1511.8, 792.4)
(727.8, 5945.9) (3173, 8394.2) AUC.sub.0-.infin. (ng h/mL) N 20 20
18 12 9 Mean 818.1 1548.2 2503.4 3336.2 5132.5 (SD) (194.6) (396.3)
(723.3) (1212.7) (1464.5) % CV 23.8 25.6 28.9 36.3 28.5 Median
793.5 1468.2 2318 3482 5006.4 (Min, Max) (622.2, 1446.7) (1166.6,
3004.3) (1551.7, 4842.7) (774.5, 6014.7) (3232.7, 8605) t.sub.1/2,
hr N 20 20 18 12 9 Mean 11.3 11.1 10.9 11.3 12.4 (SD) (2.4) (2)
(2.1) (2) (2.3) % CV 21 18.5 19.6 17.5 18.9 Median 10.8 10.6 10.6
11.6 11.7 (Min, Max) (7.6, 16) (8, 15.1 ) (8.3, 15.7) (7.8, 14.1 )
(10.4, 17.9) .sup.aAUC.sub.0-t is defined as the area under the
plasma concentration-time curve from time zero to the last sampling
time at which plasma concentrations were measurable
[0482] Dose Proportionality
[0483] Systemic exposure (AUC.sub.0-.infin.) to d-amphetamine
increased in a dose proportional manner over the range of 50-250
mg. Analysis of the linearity (table 79) of the AUC.sub.0-.infin.
vs. dose curve yielded regression parameters that confirm dose
proportionality. The slope deviated slightly from linearity due to
the presence of outliers at the two higher doses.
TABLE-US-00084 TABLE 79 Dose Proportionality of plasma
d-amphetamine AUC.sub.0-.infin. after Oral Administration of
l-lysine-d-amphetamine AUC.sub.0-.infin. (hr * ng/mL)
Regression.sup.a Slope (SE) 1.1 (0.058) (90% CI, ln scale) (0.962,
1.16) .sup.aAssessment of dose proportionality based on the power
model: log (ln AUC.sub.0-.infin.) = a + b * log (dose); where "a"
is the intercept and "b" is the slope.
[0484] The maximum observed plasma d-amphetamine concentration
(C.sub.max) was also dose proportional over the dose range of
50-250 mg. Analysis of the linearity of the C.sub.max vs. dose
curve (Table 80) yielded regression parameters that confirmed a
linear relationship of C.sub.max with dose.
TABLE-US-00085 TABLE 80 Dose Proportionality of plasma
d-amphetamine for C.sub.max after Oral Administration of
Lisamfetamine Dimesylate C.sub.max (ng/mL) Regression.sup.a Slope
(SE) 1.0 (0.054) (90% CI, ln scale) (0.891, 1.07) .sup.aAssessment
of dose proportionality based on the power model: log (ln
C.sub.max) = a + b* log (dose); where "a" is the intercept and "b"
is the slope.
[0485] Intra- and Inter-Subject Variability
[0486] Within-subject (intra-subject) and between-subject
(inter-subject, or patient to patient) variability in d-amphetamine
AUC.sub.0-.infin. and C.sub.max are presented for all doses (Table
81); and for only the 50, 100 and 150 mg dose groups (Table 82) are
presented in tables 81 and 82, below.
[0487] For AUC.sub.0-.infin., when data from all dosing periods was
considered, total variability was about 28%. Within-subject and
between subject variability were similar (about 20%). When data
from the three lowest (50, 100, 150 mg) dosing periods was
considered (Table 82), total variability in AUC.sub.0-.infin. was
reduced (22%) and the majority of the variability was
between-subject. The within-subject variability was about half that
of the between-subject variability (10% vs. 20%). For C.sub.max,
when data from all dosing periods was considered, total variability
was about 27%. Between-subject was less than within-subject
variability (16% vs. 22%). When data from the three lowest (50,
100, 150 mg) dosing periods was considered, total variability in
C.sub.max was reduced (20%) and the majority of the variability was
between-subject. The within-subject variability was about half that
of the between-subject variability (9% vs. 18%).
TABLE-US-00086 TABLE 81 Within- and between-subject variability for
d-amphetamine - all lysine-d- amphetamine dimesylate dimesylate
doses Within-Subject Between-Subject Total Parameter Estimate 95%
CI Estimate 95% CI Estimate 95% CI Log AUC.sub.0-.infin. 0.195
(0.164, 0.240) 0.204 (0.139, 0.316) 0.282 (0.238, 0.374) Log
C.sub.max 0.215 (0.181, 0.264) 0.163 (0.096, 0.264) 0.269 (0.232,
0.346) Note: Variability is on the SD (standard deviation) scale
for log-transformed parameters.
TABLE-US-00087 TABLE 82 Within- and between-subject variability for
d-amphetamine - 50, 100 and 150 mg - lysine-d-amphetamine
dimesylate dimesylate doses Within-Subject Between-Subject Total
Parameter Estimate 95% CI Estimate 95% CI Estimate 95% CI Log
AUC.sub.0-.infin. 0.104 (0.085, 0.135) 0.197 (0.143, 0.294) 0.222
(0.177, 0.313) Log C.sub.max 0.092 (0.074, 0.119) 0.176 (0.129,
0.263) 0.198 (0.158, 0.279) Note: Variability is on the SD
(standard deviation) scale for log-transformed parameters.
Pharmacokinetic Conclusions
[0488] Plasma d-amphetamine vs. time curves for doses from 50-250
mg were similar in shape and demonstrated well-behaved, consistent,
dose-proportional behavior.
[0489] Pharmacokinetic parameters for d-amphetamine were also
dose-proportional (AUC.sub.0-.infin., C.sub.max) consistent
(t.sub.max, t.sub.1/2), and well-behaved over the whole dose range
of 50 mg-250 mg. In this study, systemic exposure, maximum plasma
concentration, time to maximum concentration and the elimination
half life were reliably predictable even at supratherapeutic doses
(more than 3.5 times the current maximum daily dose).
[0490] Intra-subject variability in d-amphetamine AUC.sub.0-.infin.
and C.sub.max was low, particularly when the three lowest doses
(50-150 mg) were evaluated. For AUC.sub.0-.infin., in the 50-150 mg
dose group, the majority of the total variability (22%) arose from
between-subject variation (20%). Intra-subject variability (10%)
was approximately half the inter-subject variability.
[0491] It will be understood that the specific embodiments of the
invention shown and described herein are exemplary only. Numerous
variations, changes, substitutions and equivalents will occur to
those skilled in the art without departing from the spirit and
scope of the invention. In particular, the terms used in this
application should be read broadly in light of similar terms used
in the related applications. Accordingly, it is intended that all
subject matter described herein and shown in the accompanying
drawings be regarded as illustrative only and not in a limiting
sense and that the scope of the invention be solely determined by
the appended claims.
* * * * *